Methods of classifying periorbital dyschromia and systems therefor

ABSTRACT

A method and system for classifying different types of periorbital dyschromia is disclosed. The method includes providing at least one of a predetermined imaging characteristic and a biological characteristic for each of three different types of periorbital dyschromia, and then measuring the appropriate characteristic on a person exhibiting periorbital dyschromia. The type of periorbital dyschromia exhibited by a person can then be determined by comparing the measured value to the predetermined value and selecting the corresponding type of periorbital dyschromia.

FIELD OF THE INVENTION

The present application is directed, generally, to methods and systems for classifying periorbital dyschromia on a person. More specifically, the present application discloses methods and systems for classifying different types of periorbital dyschromia based on visual characteristics, histology, the presence, absence and/or abundance of biomarkers and genetic indicators.

BACKGROUND OF THE INVENTION

A person's eyes are a prominent and noticeable facial feature. Thus, any desirable or undesirable aesthetic features associated with the eyes may influence an individual's perception of herself or himself or the impression that the individual makes on others. Undesirable aesthetic features may include lines, wrinkles and discoloration of the skin. For example, some people may find periorbital dyschromia, sometimes referred to as dark circles or under-eye dark circles, to be aesthetically undesirable and/or they may associate the appearance of periorbital dyschromia with fatigue and/or age. It should come as no surprise then that throughout history a variety of ways to accentuate and/or beautify the eyes have been devised. A common approach to improve the appearance of periorbital dyschromia is to use a cosmetic composition such as a concealer or the like to hide the discoloration. Using make up to hide a perceived flaw may provide a temporary cosmetic benefit, but most conventional make up products require daily application and, in some instances, may even require reapplication throughout the day. Thus, a more permanent solution is desired to reduce and/or eliminate some of the undesirable aesthetic features commonly found around the eye.

As mentioned above, a particularly undesirable aesthetic eye feature is periorbital dyschromia. In an effort to find a solution to the problem of periorbital dyschromia, researchers have previously tried to identify its underlying causes. One theory suggests that periorbital dyschromia occurrence is based on the difference between the thickness of the skin in the periorbital region (i.e., the skin around the eye) and the thickness of the skin in other areas of the face such as the cheeks. It is known that the skin surrounding the eyes is typically thinner than other facial skin (e.g., on the order of 0.33 to 0.36 ram or 3 to 5 times thinner than the rest of the skin of the face). As a result, some conventional theories posit that the skin around the eyes may be easily dehydrated and particularly vulnerable to adverse impact of external factors such as heat, stress, tobacco, UV rays, and excessive facial expressions. According to these theories, the skin tissues around the eye area may undergo multiple variations throughout the day, such as vascularization, hydration and turgescence, which contributes to swelling/puffiness and/or the appearance of dark circles under the eyes. While the thinness of the skin in the periorbital region may play a role in the occurrence of periorbital dyschromia, it is not a suitable characteristic for distinguishing different types of periorbital dyschromia. This is because most people generally have thinner skin in the periorbital region compared to other areas of the face, yet not all people have the same type of periorbital dyschromia or, in sonic instances, any periorbital dyschromia at all. Thus, there remains a need to find suitable characteristics for distinguishing different types of dark circles.

Another theory attributes the occurrence of periorbital dyschromia to hyperpigmentation, or the overproduction of melanin in the skin under the eye. But recent studies have revealed that the cause of under-eye dark circles is more often the result of a combination of factors such as deep vascular congestion/superficial vascularity, hyperpigmentation, skin translucency, and structural shadowing, alone or in combination. The belief that a variety of factors are responsible for causing periorbital dyschromia has led to attempts to classify periorbital dyschromia into discrete types according to the different underlying factor(s) believed to be responsible for the discoloration. But these attempts have failed to provide a commercially viable method of classifying periorbital dyschromia or a system that is suitable for developing and marketing cosmetic products that target periorbital dyschromia. In some instances, there were too many classes of periorbital dyschromia, which requires a manufacturer to develop more cosmetic products or line ups and/or market a more complex treatment method. In some instances, the system used to classify the types of periorbital dyschromia may be too complex to allow consumer to self-diagnose, for example, at home or in a commercial environment such as a retail store.

Another theory suggests that chromophores such as melanin and hemoglobin present in the undereye region contribute to the occurrence of periorbital dyschromia. Melanin is a naturally produced pigment in the skin, and hemoglobin is the iron-containing oxygen-transport metalloprotein in the red blood cells of all humans. It is commonly believed that melanin and hemoglobin are primarily responsible for the skin tone of a person. Thus, some recent attempts to classify dark circles have included analyzing relatively small areas of skin on the lower eyelid with a device that correlates certain wavelengths of reflected light into measurements of melanin and/or hemoglobin present in the skin. While this approach may be suitable for providing an indication of the melanin and/or hemoglobin content of the particular area of skin analyzed, it has several drawbacks. For example, the amount of melanin and/or hemoglobin present in the skin or in the blood vessels in under-eye skin can vary based on a wide variety of environment and/or biological influences, which current analytical approaches do not consider. Additionally, current approaches to classifying dark circles based on melanin and hemoglobin levels may not take into account one or more of a variety of other factors believed to contribute to periorbital dyschromia, which are important to include in a robust classification method and/or system suitable for use in commerce. Previous attempts to analyze periorbital dyschromia based on melanin and hemoglobin measurements also focused on the skin of the lower eyelid. Since it is not uncommon for periorbital dyschromia to occur in the periorbital region above the eye and in portions of the undereye region, but not the entire undereye region, it is important to holistically analyze periorbital dyschromia in the periorbital region.

It is currently recognized that periorbital dyschromia is a multifactorial pathogenesis that is not well elucidated. Past attempts to classify periorbital dyschromia may have shown that there are different types of periorbital dyschronia, but they still have failed to provide a suitable method or system of classifying the different types. Further, there is a need for products, product line ups and/or treatment regimens that are particularly suited for treating different types of periorbital dyschromia. For example, some researchers may have recognized that there are different types of periorbital dyschromia, but they still propose treating different types of periorbital dyschromia with the same composition or material. Other researchers have suggested that a “one size fits all” approach to treating periorbital dyschromia may not be suitable, due to the numerous and not well understood differences in the underlying causes of periorbital dyschromia, but they do not proffer a suitable alternative. Thus, there remains a need for a suitable method and system of classifying periorbital dyschromia, and, in particular, one that is more suitable for commercial use. Otherwise, manufacturers may find it too difficult and/or unpredictable to design and market products and treatment regimens for the diverse array of consumers who typically use their products. Similarly, in some instances, it may be desirable for consumers to self-diagnose, and thus it would be desirable for the method and system to be user friendly, for example, in a commercial setting such as a retail store.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

FIG. 1 is an illustration of various portions of a human face.

FIGS. 2A and 2B illustrate examples of the portion of the periorbital region affected by Type I periorbital dyschromia.

FIGS. 3A and 3B illustrate examples of portion of the periorbital region affected by Type II periorbital dyschromia.

FIGS. 4A and 4B illustrate examples of portion of the periorbital region affected by Type III periorbital dyschromia.

FIGS. 5A, 6A and 7A are images of a test subject captured with a digital camera.

FIGS. 5B, 6B and 7B are Brown channel images of a test subject.

FIGS. 5C, 6C and 7C are Red channel images of a test subject.

FIGS. 5D, 6D and 7D illustrate the location of the periorbital dyschromia seen in FIGS. 5B, 6B and 7B, respectively.

FIGS. 5E, 6E and 7E illustrate the location of the periorbital dyschromia seen in FIGS. 5C, 6C and 7C, respectively.

FIG. 8 is a 20× micrograph of a skin tissue sample from a subject exhibiting No Dyschromia.

FIG. 9 is a 20× micrograph of a skin tissue sample from a subject exhibiting Type I periorbital dyschromia.

FIG. 10 is a 20× micrograph of a skin tissue sample from a subject exhibiting Type II periorbital dyschromia.

FIG. 11 is a 20× micrograph of a skin tissue sample from a subject exhibiting Type III periorbital dyschromia.

FIG. 12 illustrates the Fontana-Masson positive bodies observed per field of view for each type of periorbital dyschromia and a No Dyschromia condition.

FIG. 13 illustrates the normalized PTCA amount measured for each type of periorbital dyschromia and a No Dyschromia condition.

FIG. 14 illustrates an example of a masked region corresponding to Zone 1.

FIG. 15 illustrates an example of a masked region corresponding to Zone 2.

FIG. 16 illustrates an example of a masked region corresponding to Zone 4.

FIG. 17 illustrates an example of a masked region corresponding to the cheek.

FIG. 18 illustrates an example of a masked region corresponding to the cheek.

FIG. 19 shows a comparison of the R values for different types of periorbital dyschromia in Zone 1.

FIG. 20 is a comparison of the B values for different types of periorbital dyschromia in Zone 1.

FIG. 21 is a comparison of the G values for different types of periorbital dyschromia in Zone 1.

FIG. 22 is a comparison of the B/G ratios for different types of periorbital dyschromia in Zone 1.

FIG. 23 is a comparison of the R values for different types of periorbital dyschromia in Zone 2.

FIG. 24 is a comparison of the G values for different types of periorbital dyschromia in Zone 2

FIG. 25 is a comparison of the B values for different types of periorbital dyschromia in Zone 2.

FIG. 26 is a comparison of the B/G ratios for different types of periorbital dyschromia in Zone 2.

FIG. 27 is a comparison of the R values for different types of periorbital dyschromia in Zone 4 (i.e., Zones 1 and 2, in combination).

FIG. 28 is a comparison of the G values for different types of periorbital dyschromia in Zone 4 (i.e., Zones 1 and 2, in combination).

FIG. 29 is a comparison of the B values for different types of periorbital dyschromia in Zone 4 (i.e., Zones 1 and 2, in combination).

FIG. 30 is a comparison of the B/G ratios for different types of periorbital dyschromia in Zone 4 (i.e., Zones 1 and 2, in combination).

SUMMARY OF THE INVENTION

In order to provide a solution to the aforementioned problems, disclosed herein is a method of classifying periorbital dyschromia comprising providing a first predetermined imaging value for a first type of periorbital dyschromia, a second predetermined imaging value for a second type of periorbital dyschromia, and a third predetermined imaging value for a third type of periorbital dyschromia. The method includes indentifying skin in a periorbital region of a person comprising periorbital dyschromia and measuring an imaging value of at least a portion of the identified skin. The measured imaging value is compared to the predetermined imaging values, and the person is classified as having the first type of periorbital dyschromia when the measured image value corresponds to the first predetermined imaging value, as having the second type of periorbital dyschromia when the measured imaging value corresponds to the second predetermined imaging value, or as having the third type of periorbital dyschromia when the measured imaging value corresponds to the third predetermined imaging value.

DETAILED DESCRIPTION OF THE INVENTION

All percentages are by weight of the personal-care composition, unless otherwise specified. All ratios are weight ratios, unless specifically stated otherwise. All numeric ranges are inclusive of narrower ranges; delineated upper and lower range limits are interchangeable to create further ranges not explicitly delineated. The number of significant digits conveys neither limitation on the indicated amounts nor on the accuracy of the measurements. All measurements are understood to be made at about 25° C. and at ambient conditions, where “ambient conditions” means conditions under about one atmosphere of pressure and at about 50% relative humidity.

Definitions

“Connectivity map” and “C-map” refer broadly to devices, systems, articles of manufacture, and methodologies for identifying relationships between cellular phenotypes or cosmetic conditions, gene expression, and perturbagens, such as cosmetic actives. A description of connectivity mapping and methods of using connectivity mapping to identify genes and/or compositions of interest can be found in U.S. Publication No. 2012/0283112 titled “Systems and Methods For Identifying Cosmetic Agents For Skin Care Compositions” filed by Binder, et al., on Feb. 22, 2012 and U.S. Publication Nos. 2013/0259816, 2013/0261006, 2013/0261024 and 2013/0261007 all filed by Hakozaki, et al., on Mar. 27, 2013.

“Cosmetic” means providing a desired visual effect on an area of the human body. The visual cosmetic effect may be temporary, semi-permanent or permanent. Some non-limiting examples of “cosmetic products” include products that leave color on the face, such as foundation, mascara, concealers, eye liners, brow colors, eye shadows, blushers, lip sticks, lip balms, face powders, solid emulsion compact, and the like.

“Cosmetic agent” means any substance, as well any component thereof, intended to be rubbed, poured, sprinkled, sprayed, introduced into, or otherwise applied to a mammalian body or any part thereof. Cosmetic agents may include substances that are Generally Recognized as Safe (GRAS) by the US Food and Drug Administration, food additives, and materials used in non-cosmetic consumer products including over-the-counter medications. In some embodiments, cosmetic agents may be incorporated in a cosmetic composition comprising a dermatologically acceptable carrier suitable for topical application to skin. A cosmetic agent includes, but is not limited to, (i) chemicals, compounds, small or large molecules, extracts, formulations, or combinations thereof that are known to induce or cause at least one effect (positive or negative) on skin tissue; (ii) chemicals, compounds, small molecules, extracts, formulations, or combinations thereof that are known to induce or cause at least one effect (positive or negative) on skin tissue and are discovered, using the provided methods and systems, to induce or cause at least one previously unknown effect (positive or negative) on the skin tissue; and (iii) chemicals, compounds, small molecules, extracts, formulations, or combinations thereof that are not known have an effect on skin tissue and are discovered, using the provided methods and systems, to induce or cause an effect on skin tissue.

Some examples of cosmetic agents or cosmetically actionable materials can be found in: the PubChem database associated with the National Institutes of Health, USA; the Ingredient Database of the Personal Care Products Council; and the 2010 International Cosmetic Ingredient Dictionary and Handbook, 13^(th) Edition, published by The Personal Care Products Council; the EU Cosmetic Ingredients and Substances list; the Japan Cosmetic Ingredients List; the Personal Care Products Council, the SkinDeep database; the FDA Approved Excipients List; the FDA OTC List; the Japan Quasi Drug List; the US FDA Everything Added to Food database; EU Food Additive list; Japan Existing Food Additives, Flavor GRAS list; US FDA Select Committee on GRAS Substances; US Household Products Database; the Global New Products Database (GNPD) Personal Care, Health Care, Food/Drink/Pet and Household database; and from suppliers of cosmetic ingredients and botanicals.

Other non-limiting examples of cosmetic agents include botanicals (which may be derived from one or more of a root, stem bark, leaf, seed or fruit of a plant). Some botanicals may be extracted from a plant biomass (e.g., root, stem, bark, leaf, etc.) using one more solvents. Botanicals may comprise a complex mixture of compounds and lack a distinct active ingredient. Another category of cosmetic agents are vitamin compounds and derivatives and combinations thereof, such as a vitamin B3 compound, a vitamin B5 compound, a vitamin B6 compound, a vitamin B9 compound, a vitamin A compound, a vitamin C compound, a vitamin E compound, and derivatives and combinations thereof (e.g., retinol, retinyl esters, niacinamide, folic acid, panthenol, ascorbic acid, tocopherol, and tocopherol acetate). Other non-limiting examples of cosmetic agents include sugar amines, phytosterols, hexamidine, hydroxy acids, ceramides, amino acids, and polyols.

“Data architecture” refers generally to one or more digital data structures comprising an organized collection of data. In some embodiments, the digital data structures can be stored as a digital file (e.g., a spreadsheet file, a text file, a word processing file, a database file, etc.) on a computer readable medium. In some embodiments, the data architecture is provided in the form of a database that may be managed by a database management system (DBMS) that is be used to access, organize, and select data (e.g., instances and gene expression signatures) stored in a database.

“Disposed” refers to an element being located in a particular place or position relative to another element.

“Effective amount” means an amount of a compound or composition sufficient to significantly induce a positive or desired benefit, (e.g., a positive skin or feel benefit, reverse the expression of a gene, group of genes and/or gene signature), including independently or in combinations the benefits disclosed herein, but low enough to avoid serious side effects, i.e., to provide a reasonable benefit to risk ratio, within the scope of sound judgment of the skilled artisan.

“Gene expression signature” refers to a rationally derived list, or plurality of lists, of genes representative of a skin tissue condition or a skin agent. In specific contexts, the skin agent may be a benchmark skin agent or a potential skin agent. Thus, the gene expression signature may serve as a proxy for a phenotype of interest for skin tissue. A gene expression signature may comprise genes whose expression, relative to a normal or control state, is increased (up-regulated), whose expression is decreased (down-regulated), and combinations thereof. Generally, a gene expression signature for a modified cellular phenotype may be described as a set of genes differentially expressed in the modified cellular phenotype over the unmodified cellular phenotype. A gene expression signature can be derived from various sources of data, including but not limited to, from in vitro testing, in vivo testing and combinations thereof. In some embodiments, a gene expression signature may comprise a first list representative of a plurality of up-regulated genes of the condition of interest and a second list representative of a plurality of down-regulated genes of the condition of interest.

“Gene expression profiling” refers to the measurement of the expression of multiple genes in a biological sample using any suitable profiling technology. For example, the mRNA expression of thousands of genes may be determined using microarray techniques. Other emerging technologies that may be used include RNA-Seq or whole transcriptome sequencing using NextGen sequencing techniques. Gene expression profiling may be used to generate a gene expression signature.

“Joined” means configurations whereby an element is directly secured to another element by affixing the element directly to the other element, and configurations whereby an element is indirectly secured to another element by affixing the element to intermediate member(s) that in turn are affixed to the other element.

“Keratinous tissue,” means keratin-containing tissue layers disposed as the outermost protective covering of mammals which includes, but is not limited to, skin, hair, and nails.’

“Lateral” means a direction that is generally perpendicular to an imaginary centerline that bisects the human body into right and left mirror images. Directions that are within forty-five degrees of being perpendicular to the imaginary centerline are considered lateral.

“Longitudinal” means a direction that is generally parallel to an imaginary centerline that Bisects the human body into right and left mirror images. Directions that are within forty-five degrees of being parallel to the imaginary centerline are considered longitudinal.

“Microarray” refers broadly to any ordered array of nucleic acids, oligonucleotides, proteins, small molecules, large molecules, and/or combinations thereof on a substrate that enables gene expression profiling of a biological sample. Non-limiting examples of microarrays are available from Affymetrix, Inc.; Agilent Technologies, Inc.; Illumina, Inc.; GE Healthcare, Inc.; Applied Biosystems, Inc.; Beckman Coulter, Inc.; etc.

“Periorbital” means around the orbit of the eye. The periorbital region of a person is the area of the face generally disposed around the eye socket and typically lies between the bottom of the brow and the top of the cheek in the longitudinal direction and between the bridge of the nose and the temple in the lateral direction.

“Periorbital dyschromia” is a condition that occurs when the tone of skin in the periorbital region of person is noticeably different from the tone of skin in a nearby portion of the face, such as the cheek, nose, forehead, temple and/or another portion of the periorbital region. Perioribital dyschromia is bilateral, (i.e., it occurs in the periorbital region of both sides of the face). Periorbital dyschromia may appear as a result of hyperpigmented and/or hypopigmented skin disposed in the periorbital region. Periorbital dyschromia may be identified and/or classified according to one or more of the indicators described in more detail below. Periorbital dyschromia herein is classified into one of three types (i.e., Type I, Type II or Type III). The three types of periorbital dyschromia are described and defined in more detail below, and can be readily determined in accordance with the methods herein.

“Personal care composition” means a cosmetic composition or a skin care composition suitable. Is it to be appreciated that a personal care composition may provide both a cosmetic benefit and a skin health benefit,

“Reverse,” when referring to the gene expression of a gene, means that the expression of the gene is changed such that it is opposite of the expression indicated in a gene signature in a significant way (e.g., p-value <0.1, p-value <0.05, p-value <0.01, p-value <0.001, or p-value <0.0001 as determined by a statistical test like ANOVA or to a t-test). For example, if a gene expression signature indicates that a particular gene is up-regulated, then reversing the expression of the gene can mean that the gene is down-regulated relative to the indicated gene expression signature with a p-value of less than 0.05 as determined by a statistical test like ANOVA or t-test. When referring to gene expression signatures, the term “reversing” depends on the method used to determine the change in gene expression signature. For example, when using connectivity mapping, a connectivity score is generated to represent an amount of differential expression relative to a known gene expression signature, e.g., stored in a data architecture, and the connectivity score can be used as a measure of the amount of reversal in a gene expression signature in a significant way (e.g., p-value <0.1, p-value <0.05, p-value <0.01, p-value <0.001, or p-value <0.0001.

“Skin” means the outermost protective covering of mammals that is composed of cells such as keratinocytes, fibroblasts and melanocytes. Skin includes an outer epidermal layer and an underlying dermal layer. Skin may also include hair and nails as well as other types of cells commonly associated with skin, such as, for example, myocytes, Merkel cells, Langerhans cells, macrophages, stem cells, sebocytes, nerve cells and adipocytes.

“Skin care” means regulating and/or improving skin condition. Some nonlimiting examples of skin care benefits include improving skin appearance and/or feel by providing a smoother, more even appearance and/or feel; increasing the thickness of one or more layers of the skin; improving the elasticity or resiliency of the skin; improving the firmness of the skin; and reducing the oily, shiny, and/or dull appearance of skin, improving the hydration status or moisturization of the skin, improving the appearance of fine lines and/or wrinkles, improving skin texture or smoothness, improving skin exfoliation or desquamation, plumping the skin, improving skin barrier properties, improve skin tone, reducing the appearance of redness or skin blotches, and/or improving the brightness, radiancy, or translucency of skin. Some nonlimiting examples of “skin care products” include skin creams, moisturizers, lotions, and body washes.

“Skin-care active” means a compound or combination of compounds that, when applied to skin, provide an acute and/or chronic benefit to skin or a type of cell commonly found therein. Skin-care actives may regulate and/or improve skin or its associated cells (e.g., improve skin elasticity; improve skin hydration; improve skin condition; and improve cell metabolism).

“Skin-care composition” means a composition that regulates and/or improves skin condition.

“Skin tone” refers to the perceived color or pigmentation of skin, especially with regard to the evenness of the coloration or pigmentation. “Skin tone” may also include other characteristics of skin that contribute to a consumer perception of overall tone. For example, pore size and distribution, and skin texture may also be considered attributes of overall skin tone.

“Software” and “software application” mean one or more computer readable and/or executable instructions that cause a computing device or other electronic device to perform functions, actions, and/or behave in a desired manner. The instructions may be embodied in one or more various forms like routines, algorithms, modules, libraries, methods, and/or programs. Software may be implemented in a variety of executable and/or loadable forms and can be located in one computer component and/or distributed between two or more communicating, co-operating, and/or parallel processing computer components and thus can be loaded and/or executed in serial, parallel, and other manners. Software can be stored on one or more computer readable medium and may implement, in whole or part, the methods and functionalities of the present invention.

“Topical application” means to apply or spread the compositions of the present invention onto the surface of the keratinous tissue.

Before now, the underlying causes of periorbital dyschromia were not particularly well elucidated. However, it has unexpectedly been found that there are common themes associated with periorbital dyschromia that can lend themselves to differentiation based on a variety of relatively straightforward evaluation techniques. Previous attempts to classify periorbital dyschromia did not appreciate that periorbital dyschromia can be grouped into distinct categories based on the visual evaluation, imaging, biomarker, histology, and/or genetic analysis techniques disclosed herein. Based on these newly discovered distinctions, it is believed that the methods and systems described herein, individually and/or collectively, may enable the development and/or marketing of improved cosmetic products and/or treatment regimens particularly suited for treating different types of periorbital dyschromia.

Visual Evaluation

Periorbital dyschromia may be classified into three different types as well as a “No Dyschromia” condition using visual evaluation techniques. In particular, the location and/or tone(s) of a dyschromic portion of periorbital skin may be analyzed visually to determine whether perirorbital dyschromia is present and, if present, which type is present. For example, a No Dyschromia condition may be visually characterized by the lack of an uneven or discontinuous skin tone in the periorbital region. Visual evaluation may be done by an expert grader (i.e., someone trained to visually classify periorbital dyschromia), for example, as described in more detail in the Methods section below, or the visual classification may be done by a non-expert (e.g., a consumer who self-diagnoses) based on, for example, a set of instructions or a visual cue.

FIG. 1 illustrates the periorbital region 10 of a human face 5 divided into three zones. Zone 1 11 is disposed generally under the eye in the longitudinal direction (represented by the arrow Y) and extends under the eye in the lateral direction (represented by the arrow X) from the inner canthus 4 (i.e., the corner of the eye proximate the nose 20) to an area under the eye that is less than the complete distance to the outer canthus 6 (i.e., the corner of the eye spaced furthest from the nose). For example, as illustrated in FIG. 1, Zone 1 11 may extend from the inner canthus 4 to about the middle of the undereye portion of the periorbital region 10. But it is to be appreciated that, in some individuals, Zone 1 may extend more than halfway or less than halfway across the under-eye region, but typically not more than 90% of the distance from the inner canthus 4 to the outer canthus 6. Zone 2 12 extends from the distal edge of Zone 1 11 (i.e., the portion of Zone 1 furthest from the nose 20 in the lateral direction) to the outer canthus 6 of the eye. Zone 3 is disposed above the eye and extends laterally from the inner canthus 4 to the outer canthus 6. Zone 3 13 also extends generally in the longitudinal direction Y from the upper eyelid to the bottom of the eyebrow. In some embodiments, it may be desirable to use a target portion of skin disposed on the cheek 7 as a reference or control when evaluating the location and/or tones of periorbital dyschromia.

“Type I” periorbital dyschromia may be visually characterized by continuous discoloration of both the upper and lower eyelid skin. The dyschromic periorbital skin associated with Type I periorbital dyschromia typically includes substantially uniform brown, yellow and/or orange tones, which may resemble the color of tanned skin or an age spot. Additionally or alternatively, Type I periorbital dyschromia may be visually characterized by being generally present in Zones 1, 2 and 3 of the periorbital region of a person.

“Type II” may be visually characterized by continuous discoloration of the lower eyelid skin. The discolored periorbital skin associated with Type II periorbital dyschromia typically includes substantially uniform purple, pink and/or bluish tones, which may resemble bruised skin. Additionally or alternatively, Type II periorbital dyschromia may be visually characterized by being generally present in Zone 1 of the periorbital region and generally absent from Zone 2 and Zone 3.

“Type III periorbital dyschromia” typically includes a combination of characteristics of Types I and II, such as color and location in the periorbital region. In some instances, Type III perioribital dyschromia may be characterized by discontinuous discoloration of the undereye portion of the periorbital region, which is different from the more uniform discoloration associated with Types I and II. The patches of discolored periorbital skin associated with Type III periorbital dyschromia are generally present in Zone 1 and Zone 2 and may also be present in the inner portion of Zone 3 (i.e., the portion of Zone 3 that is closest to the nose).

FIGS. 2A and 2B illustrate examples of Type I periorbital dyschromia, which is represented by the shaded portions 200 and 201, respectively, of the periorbital region. FIGS. 3A and 3B illustrate examples of Type II periorbital dyschromia (i.e., the shaded portions 300 and 301, respectively, of the periorbital region). FIGS. 4A and 4B illustrate examples of Type III periorbital dyschromia (i.e., the shaded portions 400 and 401, respectively, of the periorbital region).

Imaging Classification—RGB

Type I, Type II and Type III periorbital dyschromia may be distinguished from one another using known imaging technique such as RGB and/or RBX® color imaging and/or gray scale imaging. An image of the region of interest (“ROI”) (e.g., periorbital region of a person) is captured by an image capture device, and at least a portion of the image is analyzed (e.g., by a computer) and assigned an imaging value based on the analysis. This determined imaging value may then be compared to a predetermined imaging value, which can be a single value or a range of values (e.g., one or more of the RGB values disclosed herein). The predetermined imaging value defines a particular type of periorbital dyschromia, and the comparison enables a user to identify which type of periorbital dyschromia, if any, is exhibited by the person. For example, RGB color imaging uses an additive color model wherein red, green, and blue light are added together in various ways to reproduce a broad array of colors. The RGB color model is particularly suited for sensing, representing, and displaying images in electronic systems such as televisions and computers. RGB is a device-dependent color model, which means that different devices may detect or reproduce a given RGB value differently, since the color elements (such as phosphors or dyes) and their response to the individual R, G, and B levels may vary from manufacturer to manufacturer, or even in the same device over time. Thus, it can be important to use color correction to ensure that an RGB value defines the same color across devices or in the same device over time. Color correction can be readily accomplished by one skilled in the art using commercially available software and techniques.

Classifying periorbital dyschromia according to the imaging classification methods and systems described herein depends on, among other things, the skin tone of an individual. For example, people who are Fitzpatrick I, II and III skin types according to the commonly known Fitzpatrick Scale, may have different color values associated with the different types of periorbital dyschromia as compared to people who are Fitzpatrick skin type IV, V or VI. In some of the examples and descriptions below, the different types of periorbital dyschromia are in relation to people who are Fitzpatrick skin type I, II and/or III (e.g., the imaging values and ratios). However, it is believed, without being limited by theory, that the classification methods and systems herein can be reapplied to people with Fitzpatrick IV, V and VI skin types in the substantially the same manner. In some instances, it may be desirable to normalize the imaging values of one or more zones in the periorbital region to a reference or control value such as an imaging value associated with the cheek. The normalized value may then be used to determine whether periorbital dyschromia is present and, if so, which type.

Type I periorbital dyschromia may be characterized by generally having lower RGB values relative to Types II and III as well as a different ratio of B value to G value (“B/G”). Type II periorbital dyschromia may be characterized by generally having higher RGB values compared to Types I and III. Type III periorbital dyschromia may include characteristics of both Type I and Type II. Tables 1, 2 and 3 below show ranges of color corrected RGB values, B/G ratios, L*a*b* values, chroma values and hue values that may be used to characterize Type I, Type II and Type III periorbital dyschromia, respectively, exhibited by a person having a Fitzpatrick skin type of I, II or III. The values provided in Tables 1, 2 and 3 correspond to a particular portion of the periorbital region referred to as a “mask.” Masks, and in particular Mask A, Mask B and Mask C are described in more detail below in the Imaging Method, which provides a suitable method of determining imaging values.

TABLE 1 Type I Imaging Value Mask A Mask B Mask C R 118-166 135-187 128-178 G  83-119  94-136  89-129 B  76-108  81-112  79-112 B/G 0.75-0.92 0.73-0.86 0.74-0.89 L 48-54 43-61 41-59 A 13-18  9-17 12-18 b  9-23 14-25 12-23 Hue (h) 35-53 42-60 39-56 Chroma (C) 16-28 17-28 17-28

TABLE 2 Type II Imaging Value Mask A Mask B Mask C R 148-176 165-201 159-186 G 102-135 128-160 115-148 B  96-128 108-145 103-136 B/G 0.86-0.98 0.82-0.93 0.84-0.94 L 48-60 57-69 52-64 a 11-20  9-19 10-19 b  7-16 10-20  9-17 Hue (h) 24-54 33-61 32-57 Chroma (C) 15-23 15-22 15-22

TABLE 3 Type III Imaging Value Mask A Mask B Mask C R 142-172 160-188 153-177 G  97-129 110-148 105-137 B  88-114  92-129  90-119 B/G 0.81-0.95 0.79-0.89 0.80-0.92 L 46-57 51-64 49-60 a 11-20 10-17 11-18 b  9-19 13-21 11-19 Hue (h) 29-54 42-62 36-57 Chroma (C) 17-24 18-24 18-24 Imaging Classification—RBX

Attempts have been made in the past to classify periorbital dyschromia based on melanin and/or hemoglobin content, which are believed to be the chromophores primarily responsible for skin tone. For example, some previous attempts used conventional instruments (e.g., Mexameter MX-18) to measure the amount of melanin and hemoglobin in the skin. The measured values are then compared to the different types of periorbital dyschromia observed in test subjects to identify any correlations that might be used in classifying the different types of periorbital dyschromia. However, these past attempts concluded that measuring melanin and hemoglobin in this manner does not provide a useful tool for differentiating periorbital dyschromia.

In contrast with previous findings, it has been found that certain imaging technology such as RBX® brand imaging technology, available from Canfield Scientific, Inc., New Jersey, may provide a suitable distinction between Type I, Type II and Type III periorbital dyschromia, based on differences in the Red Channel and Brown channel images, which can correspond to the melanin and hemoglobin present in the skin. This is an unexpected finding, since previous studies have concluded that using imaging systems to measure colorants or chomophores in the skin does not provide a suitable means for classifying periorbital dyschromia.

RBX® brand imaging technology is based on the premise that skin color is characterized by a limited number of colorants, or chromophores, within the layers of skin. In normal, healthy skin, melanin particles, which are primarily responsible for the overall skin color, are small and uniformly distributed resulting in a smooth, even skin tone. Hemoglobin occurs within the vascular structure at the papillary dermis, a sub-layer of skin, in oxygenated and deoxygenated forms and is responsible for red colorations of skin tone. However, it may not be desirable to quantify skin pigmentation by only measuring the total attenuation of broadband light because, apart from melanin, hemoglobin in skin also absorbs visible light in a wavelength dependent manner. Therefore, methods are required to measure light absorbance in more than one spectral band. RBX® brand imaging technology transforms a conventional RGB image (e.g., captured by a conventional digital camera) into a color space where the Red and Brown channels represent hemoglobin and melanin distribution, respectively.

In some instances, Type I periorbital dyschromia may be characterized by relatively high Brown and Red channel contrast in the periorbital region as compared to other portions of the face (e.g., cheek, chin, nose or forehead). FIGS. 5A to 5E illustrate an example of Type I periorbital dyschromia observed in an RBX® image. FIG. 5A is a full color image of a test subject captured with a digital camera. As can be seen in FIG. 5A, periorbital dyschromia appears in Zones I, II and III of the periorbital region of the test subject. FIG. 5B shows the Brown Channel image that corresponds to the digital image of FIG. 5A. FIG. 5C shows the Red Channel image that corresponds to the digital image from FIG. 5A. FIG. 5D provides an illustration of the location of the periorbital dyschromia provided in FIG. 5B (i.e., in the Brown channel), and FIG. 5E provides an illustration of the location of the periorbital dyschromia provided in FIG. 5C (i.e., the Red Channel). As can be seen in FIG. 5C, and as illustrated in FIG. 5E, periorbital dyschromia is present in Zones 1 and 2 and in a portion of Zone 3. However, the periorbital dyschromia appearing in Zone 3 of FIG. 5C is not present in as large an area as the periorbital dyschromia in Zone 3 of FIG. 5B. This may suggest that melanin and hemoglobin both influence the appearance of Type I periorbital dyschromia, but melanin is the more abundant chromophore. FIG. 5E provides an illustration of the periorbital dyschromia that appears in FIG. 5C (i.e., in the Red Channel).

Type II periorbital dyschromia may be characterized by relatively low Brown Channel intensity values in the periorbital skin relative to the surrounding areas of face (e.g., cheek and forehead) and high Red Channel contrast in the periorbital region as compared to other portions of the face. FIGS. 6A to 6E illustrate an example of Type II periorbital dyschromia observed in an RBX®. FIG. 6A is an image of a subject captured with a digital camera. As can be seen in FIG. 6A, periorbital dyschromia appears in Zone 1, but appears to be substantially absent from Zones 2 and 3. FIG. 6B shows the Brown channel image that corresponds to the digital image of FIG. 6A. FIG. 6C shows the Red Channel image that corresponds to the digital image of FIG. 6A. FIG. 6D provides an illustration of the location of the periorbital dyschromia provided in FIG. 6B (i.e., in the Brown channel), and FIG. 6E provides an illustration of the location of the periorbital dyschromia provided in FIG. 6C (i.e., the Red Channel). As can be seen in FIG. 6C, and as illustrated in FIG. 6E, periorbital dyschromia is visible in Zone 1, but appears to be substantially absent from Zones 2 and 3. This may suggest that melanin does not play as significant a role as hemoglobin in Type II periorbital dyschromia.

Type III periorbital dyschromia may be characterized by discontinuous patches of Brown Channel and Red channel intensity in the periorbital region. FIGS. 7A to 7E illustrate an example of Type III periorbital dyschromia observed in an RBX® image. FIG. 7A is an image of a test subject captured with a digital camera. As can be seen in FIG. 7A, periorbital dyschromia appears in Zones 1 and 3, but appears to be substantially absent from Zone 2. FIG. 7B shows the Brown channel image that corresponds to the digital image from FIG. 7A. FIG. 7C shows the Red channel image that corresponds to the digital image from FIG. 7A. FIG. 7D provides an illustration of the location of the periorbital dyschromia provided in FIG. 7B (i.e., in the Brown channel), and FIG. 7E provides an illustration of the location of the periorbital dyschromia provided in FIG. 7C (i.e., the Red Channel). As can be seen in FIG. 7B, and as illustrated in FIG. 7D, periorbital dyschromia appears in Zones 1 and 3 of the periorbital region of the test subject. And as can be seen in FIG. 7C, and as illustrated in FIG. 7E, periorbital dyschromia appears in Zone 1, but appears to be substantially absent from Zones 2 and 3.

Histology

Type I, Type II and Type III periorbital dyschromia may be distinguished from one another using histological evaluation techniques that include, for example, sectioning and staining, followed by examination under a microscope (e.g., light or electron). In particular, it has been found that the abundance and/or location of certain cellular structures (e.g., melanin) within skin biopsy samples obtained from a discolored area of periorbital skin may be used to distinguish Type I, Type II and Type III periorbital dyschromia from one another. Skin tissue samples for use herein may be obtained, sectioned and/or stained according to any suitable method in the art. The Biopsy Method described in more detail below is an example of a suitable method of sample collection, sectioning and staining. Examples of conventional stains suitable for use herein include hematoxylin and eosin stain (“H&E stain”) and Fontana-Masson stain. H&E stain is a known for use in histology and involves the application of hemalum, which is a complex formed from aluminum ions and oxidized haematoxylin, to a tissue sample. The application of hemalum colors the nuclei of the cells in the sample dark blue or purple. The nuclear staining is followed by counterstaining with an aqueous or alcoholic solution of eosin Y, which colors eosinophilic structures such as cytoplasm in various shades of red, pink and orange. Fontana-Masson stain uses ammoniacal silver nitrate to detect cellular structures in a tissue sample that are capable of reducing the silver nitrate to metallic silver, which stains the structure black.

FIGS. 8 to 11 are micrograph images at 20× magnification of H&E-stained biopsy samples from skin disposed in the periorbital region of human test subjects. The samples were collected, sectioned and stained according to the Biopsy Method described in more detail below. Each test subject was classified as exhibiting Type I, Type II, Type III or No Dyschromia by an expert grader. The skin tissue sample shown in FIG. 8 is from a test subject classified as exhibiting No Dyschromia, and thus may serve as a suitable control. The dark (i.e., purple) bodies shown in FIG. 8 are nuclei of the various cells commonly disposed in the dermis and epidermis of an individual. The lighter (i.e., red/pink) stained areas between the nuclei corresponds to cytoplasm, connective tissue and glycoproteins.

The skin tissue sample shown in FIG. 9 is from a test subject classified as exhibiting Type I periorbital dyschromia. The white arrowheads shown in FIG. 9 point to deposits, which are brown in color when seen in a color micrograph, disposed around the nuclei of some of the cells in the sample. When the tissue samples are stained with Fontana-Masson stain, the deposits appear black (“Fontana-Masson positive deposits”), which indicates that they may be melanin. This is surprising because melanin is generally not associated with the dermis and is more commonly found in the epidermis. As can be seen in FIG. 9, the Fontana-Masson positive deposits are generally disposed around the nuclei of cells that appear to be clustered together underneath the epidermal-dermal junction. The clustered cells are believed to be fibroblasts, which account for the majority of cells typically found in a healthy dermis. Healthy fibroblasts generally appear as elongated, flat cells, but the clustered cells in FIG. 9 are round when viewed under a light microscope, which indicates that they may not be healthy fibroblasts. Thus, it is believed, without being limited by theory, that the presence of these melanin deposits in the dermis may be related to an underlying biological cause of Type I perioribital dyschromia. In particular, the round clustered appearance of the fibroblasts may be associated with an inflammation response. In view of this discovery, it may be desirable to provide a method and system of classifying periorbital dyschromia based, at least in part, on dermal melanin content. It may also be desirable to configure products and regimens suited for treating Type I periorbital dyschromia to address the presence of melanin in the dermis and/or the factors that may influence the presence of melanin in the dermis.

In addition to the discovery of unexpected amounts of Fontana-Masson positive deposits in the dermis, there also appeared to be an overabundance of melanin in the epidermis of periorbital skin tissue samples taken from Type I individuals. Thus, it may be desirable to tailor products for treating Type I periorbital dyschromia to include a cosmetic active that addresses the presence of excess melanin in the epidermis.

The skin tissue sample shown in FIG. 10 is a sample from a test subject classified as exhibiting Type II periorbital dyschromia. The Fontana-Masson positive deposits that can be seen in the dermis of the Type I sample of FIG. 9 are absent from the dermis of the Type II sample in FIG. 10. Thus, the underlying biological causes of Type I and Type II periorbital dyschromia may be different from one another, which is important since products and treatments suitable for one type of periorbital dyschromia may not suitable for another type. In particular, products and treatments for Type II periorbital dyschromia may not need to address the presence of melanin in the dermis.

Examination of the skin tissue samples taken from Type II individuals also showed a lack of melanin in the epidermis. Thus, it is also important to recognize that periorbital dyschromia may be influenced not only by hyperpigmentation (i.e., too much pigmentation), as suggested by some researchers, but surprisingly appears to be influenced by hypopigmentation (i.e., not enough pigmentation) as well.

The skin tissue sample shown in FIG. 11 is a sample obtained from a test subject classified as exhibiting Type III periorbital dyschromia. As indicated by the white arrowheads in FIG. 11, there are Fontana-Masson positive deposits present around the nuclei of some cells in the dermis, but not as many as seen in the Type I sample shown in FIG. 9. It is believed, without being limited by theory, that Type III periorbital dyschromia may occur as a result of one or more factors from Type I and Type II periorbital dyschromia acting to produce periorbital dyschromia that exhibits at least some of the characteristics of both of these types. For example, the unexpected presence of melanin in the dermis may indicate that Type III periorbital dyschromia shares a common underlying cause with Type I, but less severe as evidenced by a lower amount of dermal melanin relative to Type I. Thus, products and treatments suitable for treating Type III periorbital dyschromia may need to address the underlying causes of Type I and/or Type II periorbital dyschromia, but perhaps not to the same extent. For example, a product suitable for treating Type I periorbital dyschromia that includes an active for addressing the presence of melanin in the dermis may not need to include as much of the active.

Biomarker

Type I, Type II and Type III periorbital dyschromia may be distinguished from one another by collecting and analyzing the biomarkers present in periorbital skin. In particular, it has been found that the presence, absence and/or abundance of certain molecules in the epidermis of periorbital skin can be useful in distinguishing Type I, Type II and Type III periorbital dyschromia from one another. An example of such a molecule is pyrrole-2,3,5-tricarboxylic acid (“PTCA”), which is formed as a result oxidative degradation of eumelanin. It has been found that Type I and Type III periorbital dyschromia have higher PTCA levels than Type II, and that Type I may exhibit higher PTCA levels than Type III. FIG. 13 illustrates the comparison between PTCA levels of samples obtained from a subject classified as No Dyschromia 382, a subject who classified as Type I periorbital dyschromia 384, 2) a subject classified as Type II periorbital dyschromia 386, and 3) a test subject classified as Type III periorbital dyschromia 388. As illustrated in the chart 550 of FIG. 13, the PTCA levels of the Type I 384 and Type III 388 subjects was higher than the No Dyschromia 382 and the Type II subject 386 by a statistically significant amount. In contrast, the PTCA level of the Type II subject 386 was not higher than the No Dyschromia 382 by a statistically significant amount. PTCA levels herein are determined according to the Tape Stripping method described in more detail below.

Gene Expression

Type I, Type II and Type III periorbital dyschromia may be distinguished from one another by analyzing the expression of certain genes, individually or collectively, in the dermis and/or epidermis of periorbital skin. The gene expression signature of a periorbital skin sample may be obtained by any suitable means known in the art. For example, genetic material may be obtained from a tissue sample provide by a donor (e.g., full thickness skin biopsy that exhibits a condition of interest) and subsequently processed using any suitable technology such as, for example, microarray analysis or NextGen sequencing to provide a gene expression signature. The gene expression signatures of Type I, Type II and Type III periorbital dyschromia can then be compared to one another and/or a control to identify the differences in gene expression. Example 4 below provides an example of gene expression signatures associated with each of Type I, Type II and Type III periorbital dyschromia.

It may be desirable to distinguish the different types of periorbital dyschromia based on biological themes that correspond to the expression of certain genes, combinations of genes and/or gene families. That is, a technique sometimes referred to as theme analysis may be used to identify biological or phenotypic themes associated with the gene expression data that correspond to the different types of periorbital dyschromia. Theme analysis is a statistical analysis-based method for detecting biological patterns in gene expression data. The method uses an ontology of controlled vocabulary terms developed by the Gene Ontology (“GO”) Consortium [Ashburner, M. et al. (2000) Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet, 25, 25-29] that describe the biological processes, molecular functions and cellular components associated with gene products. Analysis involves statistical comparison of a regulated list of genes and a larger reference list of all the expressed genes, to determine if genes annotated to specific GO terms are significantly enriched in the regulated list. This analysis may reveal biological patterns when multiple genes associated with a given GO term occur on the regulated list at a frequency greater than expected by chance. Such analysis may be performed using Theme Extractor proprietary software and an algorithm that calculates a p-value of each ontology term. Data may be analyzed for statistical significance, for example, by the Fisher's Exact Test. Conventional approaches and statistical methods such as, for example, Gene Set Enrichment Analysis described by Subramanian, A., et al., in “Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles,” Proc. Natl. Acad Sci U.S.A, 102, 15545-155501 (2005) are suitable for conducting theme analysis herein.

Methods and Systems for Classifying Periorbital Discoloration

In some instances, the method herein includes indentifying a target portion of skin in the periorbital region of a person, evaluating the target skin portion based on the relevant characteristics described herein and classifying the person as having Type I, Type II or Type III periorbital dyschromia, if one or more characteristics associated with the relevant type of periorbital dyschromia are met. In some instances, the method may include classifying the type of periorbital dyschromia based solely on one of the foregoing distinguishing characteristics (i.e., visual characteristics such as RGB and/or RBX values, biomarkers, histology or gene expression). In some instances, the method includes classifying periorbital dyschromia based on a combination of two or more of the foregoing characteristics (e.g., visual and gene expression characteristics, visual and biomarker characteristics, or visual and histology characteristics).

Systems suitable for classifying periorbital discoloration herein may include an image capture device, an image analysis device, a means for communicating a type of periorbital dyschromia to a user of the system and, optionally, a set of instructions for using the system. The type of periorbital dyschromia may be selected from Type I, Type II and Type III as described herein. A user may also be classified as having No Dyschromia with the present system. The system may include a camera or similar device capable of capturing digital images of sufficient quality (e.g., greater than 8 megapixels per inch) for analysis by the image analysis device. The image capture device may be configured to store and/or transfer raw digital images and JPG digital image files. In some instances, the image analysis device may be a computer in electronic communication with the image capture device. The image analysis device should be capable of receiving and analyzing a raw digital image and/or a JPG image and converting the RGB color values of each pixel in the image to a more suitable color scale such as L*a*b* and, optionally, color and/or hue values (e.g., C* and h* values). Suitable image processing software (e.g., stored directly on the image analysis device or available for use or on the internal may be used by the image analysis device to process and/or analyze various aspects of the image. In some instances, it may be desirable to normalize the pixel colors of the image analysis system to reduce variation between images captured by different devices (e.g., using known color correction techniques and/or software). An exemplary method of normalizing an image analysis device is described in more detail below. Of course, it is to be appreciated that other suitable normalization techniques known in the art may be used.

The image analysis system may be configured to classify each pixel (or a predetermined number of pixels) into one of three groups (e.g., Type I, Type II and No Dyschromia) based on the color of the pixel. The image analysis system may then calculate the percent total number of pixels in each classification and determine which type of periorbital dyschromia is present or whether any periorbital dyschromia is present, based on the percentage of pixel types from each group. After determining the type of periorbital dyschromia present, if any, the system communicates the results of the analysis to the user, for example, visually via a monitor or television, audibly via a speaker or by any other suitable communication means known in the art.

In some embodiments, it may be desirable to place the system in a commercial environment such as a retail store to enable consumers to self-diagnose. In this way, a consumer would know which type of periorbital discoloration they might have and can select the appropriate cosmetic product to treat it.

Test Methods

Visual Classification Method

This method provides a suitable means to visually evaluate and classify periorbital dyschromia using expert graders. The visual evaluation can be done in-person as well as from images (e.g., cross-polarized images).

In-Person Grading

Prior to visual evaluation, the subjects are asked to wash their face and remove any make-up, especially eye make-up. The hair of the subject is covered with a hairnet and the head and shoulders of the subject are covered with black cloth. The subjects are asked to sit between two sets of lights that illuminate each side of their face without casting shadows or causing shine in the area of the eye. The expert grader assesses the subjects from a distance of approximately 60 cm, during which time subjects are asked to relax and refrain from making any facial expressions. The expert grader evaluates area, contrast and color of the periorbital region of the subject when classifying.

Image Grading

When visually evaluating and classify periorbital dyschromia from images, it may be desirable to capture an image of the region of interest of the test subject. To capture an image, prepare the subject by covering the hair of the subject with a hairnet and covering the head and shoulders of the subject with a black cloth. All jewelry that can be seen in an image is removed. The subject is seated on the adjustable stool and rotated towards the chin rest. The operator adjusts the stool such that the subject's chin rests comfortably on the chin rest. Front view images are captured using the Cross Polarized lighting modality. The images are evaluated in a dark room using professional color calibrated LCD monitors such as LACIE brand monitors. Graders are positioned approximately 60 cm from the monitor. Attributes such as area, intensity, contrast, color and continuity are evaluated for each subject.

Imaging Method

This method provides a suitable means for capturing a reproducible and analyzable image. Any suitable image capture device along with imaging software and other associated ancillary equipment (e.g., computer and lights) may be used. A particularly suitable imaging system is the Visia-CR® brand imaging system, available from Canfield Scientific, New Jersey. The Visia® brand imaging system incorporates a Canon® brand EOS-1Ds Mk III SLR camera, which includes a CMOS sensor and provides 21.1 Mega pixel resolution (14-bit A/D converter).

Images may be collected under different lighting modalities using standard light, UV, cross-polarization, parallel-polarization or a combination of these. For example, the values and ranges described herein are reported using a (D65/2) light source. One skilled in the art will appreciate that these values can be reported at a wide range of different illuminations (D50, D75, Illuminant A, F2, F7, F11, TL84, etc. or 2 or 10 degree observer) according to well known conversion methods, and when such conversions occur, the color values will typically change accordingly. In other words, even though the actual limits and/or ranges may change based on the conditions under which the image is captured, similar relationships among the values and ranges will still be seen. For example, if the camera has lower spectral sensitivity in the red channel than the camera described herein, the R channel response may be lower and the corresponding L*a*b*C*h* (“LabCh”) color values will be different, which in this case may result in lower a* values, higher b* values, and/or higher hues. Accordingly different camera sensitivities, lightings and relevant exposures are contemplated by the method herein, and the actual limits and/or ranges disclosed herein may vary according to the particular circumstances in which the image is captured without departing from the scope of the systems and/or methods described herein.

In preparation for image capture, test subjects are required to wash their faces and wait for at least 15 minutes to let their face dry. The hair of the subject is covered with a hairnet and the head and shoulders of the subject are covered with a black cloth. All jewelry that can be seen in an image area of interest is removed. The subject is positioned such that the subject's chin is resting comfortably on the chin rest of the imaging system, and a front image of the face (as opposed to a left-side or right-side image) can be suitably captured by the image capture device. After the subject is positioned, one or more images are captured (e.g., between 1 and 24, 2 and 20 or even between 3 and 15) with the subject's eyes open. It can be important to ensure that the subject's eyes are open when the image is captured, otherwise the closed upper eyelid may cause an inaccurate pigmentation reading. The captured image(s) are processed by converting the raw image to a .jpg file format.

Next, the .jpg format image is analyzed by a computer with suitable image analysis software. In some instances, it may be desirable to analyze only a portion of the image (e.g., Zone 1, 2 and/or 3 of the periorbital region). The portion of the image to be analyzed may be “masked,” for example, as shown in FIGS. 14 to 18, using image editing software such as Photoshop® or ImageJ®. The masked region can then be isolated and analyzed as a separate image. FIG. 14 illustrates an example of a masked region corresponding to Zone 1, which is referred to herein as “Mask A.” FIG. 15 illustrates an example of a masked region corresponding to Zone 2, which is referred to herein as “Mask B.” FIG. 16 illustrates an example of a masked region corresponding to Zones 1 and 2, which is referred to herein as “Mask C.” FIGS. 17 and 18 both illustrate an example of a masked region corresponding to the cheek. It is to be appreciated that the image need not necessarily be masked for suitable analysis, and in some instances the entire image may be analyzed. In some instances, it may be desirable to reduce the size of the image, mask and/or region of interest by several pixels (e.g., between 5 and 15 pixels) around the outer edge of the image where some shadowing may occur.

In some instances, it may be desirable to convert some or all of the RGB values in the image or a portion thereof to LabCh values. The LabCh values can be calculated using a suitable RGB conversion tool at D65 Illuminant and 2 degree observer (i.e., D65/2). This conversion can be performed by software installed on the computer or a suitable conversion tool may be found online. The conversion from RGB values to LabCh values can be performed on the entire image, a portion thereof or on one or more individual pixels. The resulting LabCh values may be averaged to provide average values for the image, mask or region of interest.

In some instances, the pixels may be analyzed individually and each pixel classified as corresponding to a particular type of periorbital dyschromia based on one or more of the LabCh values. When analyzed individually, the pixels may be analyzed according to their distribution across the different types of periorbital dyschromia. An example of classifying periorbital dyschromia based on individual pixel distributions is described below in Example 2. Since color may be perceived as being relative, depending on, for example, which instruments and/or imaging system is used, it can be important to color correct the masked region for each subject using a suitable color correction technique (e.g., according to International Color Consortium standards and practices), which helps make the color determination by the system less instrument specific. In some instances, it may be desirable to normalize the color in a region of interest (e.g., a masked region) to the basal skin tone of a nearby region (e.g., cheek). For example, the basal skin tone of the cheek may be obtained by masking a region of interest in the cheek (e.g., as illustrated in FIG. 20 or 21) and converting the RGB values in the masked region to L*a*b*C*h* values as described above. The resulting basal skin tone values for the cheek may then be subtracted from the corresponding values in the region of interest to provide normalized values. Color normalization may be performed on the entire region of interest (e.g., an average value for the ROI) or on a pixel by pixel basis for some or all of the pixels in the ROI, which may be 200,000 or more pixels. The following formula provides an illustration of color normalization. DL1=Lpixel1−Lcheekave DC1=Cpixel1−Ccheekave Dh1=hpixel1−hcheek ave

-   -   Where DL, DC and Dh are the normalized L*, C*, and h* values,         respectively, of a pixel in the mask region; Lpixel, Cpixel and         hpixel are the respective L*, C* and h* values of the pixels as         determined from the image analysis; and Lcheekave, Ccheekave and         hcheekave are the respective average L*, C* and h* values         determined for the cheek.         Biopsy Method.

The biopsy method provides a suitable means for collecting, processing, storing and staining skin tissue samples. The biopsy also provides a suitable means for analyzing the skin samples for histological features associated with the different types of periorbital dyschromia. It is to be appreciated that certain portions of the method may refer to particular tools, equipment, or materials and the use thereof, but, unless indicated otherwise, one of ordinary skill in the art will recognize that equivalent tools, equipments and/or materials and the uses thereof may also be employed without departing from the scope of this method.

Sample Collection.

Skin tissue samples herein may be collected by means of a biopsy, which involves the physical removal of skin tissue from a subject. Biopsy samples collected for use herein may be removed according to any safe and aseptic biopsy method known in the art. For example, the biopsy samples may be collected from a living subject by a physician using conventional, aseptic techniques, and then flash frozen in liquid nitrogen and stored at −80° C. Eyelid biopsy sample collection involves the removal of skin tissue (down to the fatty layer) with, for example, a small elliptical surgical snip (e.g., Westcott™ brand scissors or equivalent). In at least some subjects, the elliptical shape of the incision allows for improved healing in the eyelid areas. Biopsy samples from other areas of skin on the body (e.g., cheek, forehead, arm or leg) may be obtained using, for example, a 2 mm sterile punch biopsy instrument. The size of the resulting punch biopsy sample is typically 2.0 mm in diameter and between 0.25 mm to 1.5 cm thick. In some instances, it may be desirable to suture the biopsy site.

Sample Storage

Biopsy samples should be frozen as soon as possible to minimize sample degradation Thus, it may be desirable to embed the biopsy samples in optimum cutting temperature (“OCT”) tissue freezing medium (e.g., Tissue-tek® brand OCT tissue freezing medium available from Sakura Finetek, Calif.) within 5 minutes of biopsy collection and store the samples in a suitable freezer at a temperature of −80° C. or less.

Sample Sectioning.

Frozen biopsy samples may be sectioned using a Microm HM500 brand cryostat or equivalent. The cryostat should include a microtome capable of producing multiple slices of sectioned tissue sample ranging from 5 to 300 μm in thickness. For example, 14 μm thick sectioned tissue samples are suitable for use herein. Sectioned tissue samples may be placed on slides or other support medium suitable for further analysis (e.g., viewing under a light microscope).

Sample Staining.

Hematoxylin and Eosin stain are commonly used in pathology to get an overall assessment of the tissue and determine if structural abnormalities are present. Commercially available kits such as the Shandon Rapid Chrome Frozen Section Kit available from Thermo Scientific may be used according to the manufacturer's protocol for H&E staining of the sectioned tissue samples. Other staining techniques suitable for identifying histological features in a skin tissue sample (e.g., commercially available Fontana-Masson staining kits) may also be used herein in accordance with a manufacturer's protocol and/or another suitable method known in the art.

Tape Stripping Method

This method provides a suitable means of obtaining PTCA and/or other biomarkers from biological samples collected from test subjects. It has been discovered that biological material collected in this way may be correlated to a type of periorbital dyschromia (i.e., Type I, Type II, Type III or No Dyschromia). Samples may be collected by contacting a suitable adhesive-containing substrate (“tape strip”) with a target skin surface such that biological material on and/or in the targeted skin portion adheres to the adhesive. Examples of suitable tape strips for use herein include D-Squame® brand polyacrylate ester adhesive tape (available from CuDerm; Dallas Tex.), Durapor, Sebutape™, and Tegaderm™ brand tape, duct tape (e.g., 333 Duct Tape, Nashua tape products), Scotch® brand tape (3M Scotch 810, St. Paul, Minn.), Diamond™ brand tape (The Sellotape Company; Eindhoven, the Netherlands), Sentega™ (polypropylene tape, Sentega Eiketten BV, Utrecht, The Netherlands).

The tape strips are digested with a suitable quantity and concentration of hydrogen peroxide to obtain PTCA from eumelanin. PTCA and its stable-isotope-labeled internal standard, PTCA-¹⁸O, are then isolated from the digest by a liquid-liquid extraction procedure. The extracted analyte is subjected to reverse-phase, high-performance liquid chromatographic (“HPLC”) analysis on a Waters Atlantis dC18 (3 μm 2.1×100 mm) or equivalent. Detection and quantitation of the PTCA is by tandem mass spectrometry (MS/MS) operating under multiple reaction monitoring conditions, as described in more detail below. Calibration standards are used to quantitate extracted Quality Control samples and unknown Study Specimens. The nominal range of quantitation is 0.15 to 100 ng of PTCA per sample. The concentration of PTCA is determined in the tape strip extract and then converted into total mass/tape strip by multiplying by the extraction volume. The squame scan total protein content found on the tape strips and extracts is determined and the final results are reported as total mass of analyte (ng or μg) per total weight of protein (μg) per sample. Specimen concentrations are determined by back-calculation using a weighted (1/x2) quadratic regression of a calibration curve generated from spiked matrix standards.

Sample Collection

A suitably sized tape strip (e.g., 14 mm diameter (circular) D-Squame® brand polyacrylate ester adhesive tape) is placed on a clean area of the target skin surface (e.g., periorbital region and/or cheek) and adequate pressure is applied to obtain the sample but not cause discomfort to the subject. The tape strip sample is then stored in a suitable storage container (e.g., storage tray or tube) until subjected to the digestion process.

PTCA Analysis

The tape strip sample is carefully removed from the storage container using tweezers. The tape strip sample is curled such that the adhesive-side faces inward and the non-adhesive side faces outward to reduce the possibility of contaminating the skin sample contained on the adhesive side of the tape. The curled tape strip sample is placed as deeply as possible into a 2 mL-polypropylene analysis tube in preparation for extraction of the biological material. The tape strip sample may be transferred to the analysis tube on the day of extraction or samples may be placed on storage plates and transferred to the 2 mL analysis tubes prior to extraction.

The tape strip sample is transferred from the 2 mL analysis tube to a 4 mL cryogenic Nalgene tube. A matrix blank is also prepared. The blank acts as a control and undergoes the sample process as the samples, but does not include a tape strip sample. 0.900 mL of 2 M ammonium hydroxide and 0.900 mL of 6% hydrogen peroxide are added to each of the Nalgene tubes, capped and vortexed. The tubes are placed on a rocker to incubate for three hours at 200 Mot/minute. The tubes are removed from the rocker and carefully uncapped to release any pressure that may build up during incubation. The liquid contents of each 4 mL Nalgene tube are transferred to separate 15 mL conical tubes. The tape strip is discarded and not transferred to the 15 mL conical tube. 32 μL of 10% sodium bisulfite is added to each 15 mL conical tube, capped and vortexed. Add 50 μL internal standard solution, cap and vortex. Internal standard solution is prepared as follows:

-   -   Using the appropriate Internal Standard Reference Standard         (i.e., stable, isotope-labeled PTCA ¹⁸O) weigh approximately 5         mg of Internal Standard into a glass scintillation vial and         record the weight to at least the nearest 0.01 mg. Add of the         exact amount of 10 mM ammonium acetate solution to give exactly         a 1.00 mg/mL solution of the compound according to the following         equation:

${{Volume}\mspace{11mu}({mL})} = {\frac{{Mass}\mspace{14mu}{of}\mspace{14mu}{material}\mspace{11mu}({mg})}{1.0\mspace{14mu}{{mg}/{mL}}} \times {Purity} \times {SCF}}$

-   -    Where:         -   Purity=Decimal % purity assigned to the Analytical Reference             Standard; and         -   SCF=salt correction factor.     -   Cap with a polytetraflouroethylene-lined cap and mix thoroughly         and sonicate to dissolve the compound. Store the resulting         solutions at −70 C.         2.00 mL of methyl-tert-butyl ether, 1.00 mL of 37% HCL, and 1.00         mL of saturated ammonium sulfate are added to each tube. The         tubes are capped and vortexed. Centrifuge the samples at 3,000         RPM for 15 minutes and then transfer 1.00 mL of the organic         layer of each tube to separate liquid chromatography vials. Dry         the vials down under nitrogen as is known in the art.         Reconstitute the vials with 0.250 mL of ammonium acetate, cap         and vortex. Analyze the samples by high-performance liquid         chromatography (HPLC)/mass spectrometry (MS)/mass spectrometry         (MS).

EXAMPLES Example 1

This example illustrates how different types of periorbital dyschromia can be distinguished using the Visual Classification Method. The Munsell Book of Color, Matte Collection (Munsell Color, Inc.) nomenclature is used to describe the color appearance of the different types of periorbital dyschromia or the lack thereof. The basal skin tone in this example refers to overall tone of the skin on the cheek.

1. No Dyschromia: occurs when there is no discoloration present in the periorbital region, discoloration is present but the intensity is very low (“non-severe”), or the discoloration does not extend beyond the inner corner of the eye (e.g., into Zone 1).

2. Type 1 Periorbital Dyschromia: occurs when Zone 1, Zone 2 and Zone 3 of the periorbital skin region are affected with a “tanned-like” skin tone. The tanned-like appearance found on Type I periorbital dyschromia may result from a darkening of the periorbital skin and an increase of the chroma relative to the basal skin tone. In this scenario, the hue in the periorbital skin is similar to basal skin hue. The Munsell Book of Color, Matte Collection (Munsell Color, Inc.) nomenclature can be used to describe this visual classification. For example, a Caucasian female (i.e., Fitzpatrick skin type I to III) who has basal skin tone of 10R 7/4 according to the Munsell Color nomenclature (H V/C—which corresponds to “hue”, “value” (i.e., darkness) and “chroma” respectively), will be classified as Type 1 when the periorbital skin V value is 6 or lower and C value is 5 or higher (e.g., 10R 6/5 10R6/6, 10R 5/5, 10R 5/6). In another example, a Caucasian female with basal skin tone 10R 7/4 will be classified as Type I when the periorbital skin value 2.5YR 5/5 or 2.5YR 5/6. 3. Type II Periorbital Dyschromia: occurs when Zone 1 of the periorbital skin is affected with a “bruised-liked” tone relative to the basal skin. Bruised appearance of the periorbital skin can be a result of a darkening of the skin accompanied by a decrease in chroma relative to the basal skin tone. In this case, the periorbital skin hue may be similar to basal skin hue. For example, using the Munsell Matte Color collection, a Caucasian female with a basal skin tone of 10R 7/4 may manifest a bruised appearance characteristic of Type II periorbital dyschromia when the V value for Zone I and/or II drops to 6 or below and the chroma value drops to 4 or below (e.g., 10R 6/3, 10R6/2, 10R 5/3, 10R 5/2). The bruised appearance characteristic of Type II periorbital dyschromia can also result when the periorbital skin gets darker, chroma decreases and red hues are present. A Caucasian female with a basal skin tone of 10R 7/4 will manifest bruised appearance of Type II periorbital dyschromia when the periorbital skin hue is 7.5R, V is less than 7 and Chroma is less than 4 (e.g., 7.5R 6/2, 7.5R 5/2 7.5R 6/3, or 7.5R5/3). 4. Type III Periorbital Dyschromia: may occur when elements of both Type I and Type II periorbital dyschromia are present. Type III is typically observed in Zone I and Zone 2 of the periorbital region, and may a display “bruised-like” and “tanned-like” appearance. The “bruised-like” and “tanned-like” appearances may be observed in the same or different periorbital Zones simultaneously. For example, a Caucasian female may exhibit Type III periorbital dyschromia when Zone I displays a bruised appearance and Zone II displays tanned appearance. Type III periorbital dyschromia can also occur when bruised and tanned appearance occur simultaneously in Zone I and Type III can also occur when Zone 1 and/or Zone 2 of the periorbital region are affected with “bruised-like” and “tanned like” appearance and Zone 3 is effected by either “bruised-like” or “tanned-like” appearance. Type III can also occur when there is a darkening in the periorbital skin when compared to basal skin such as the cheek with no change in chroma value. For a Caucasian female with basal skin tone 10R 7/4, Type III periorbital dyschromia may occur from a darkening in the skin with no change in chroma, can be represented, for example, by 10R 6/4, 7.5R 6/4.

Example 2

This example illustrates the distinctions between Type I, Type II and Type III periorbital dyschromia using imaging methods. Using the methods described herein, images were obtained from test subjects identified as having Type I, Type II, or Type III periorbital dyschromia, and the RGB values and B/G ratios from each of zones 1, 2 and 4 were determined for each subject. Zone 4 refers to the combination of Zones 1 and 2. The zones were masked as described in the imaging analysis method above. Zone 4 was masked as illustrated in FIG. 16. Side-by-side plots of the various RGB values and B/G ratios are illustrated in FIGS. 19 to 30. The percentile shown on the y-axis of the charts in FIGS. 19 to 30 represents the percentile ranking of each individual in the distribution. The data point at the 50^(th) percentile represents the median of the data set (i.e., the individual who was in the middle of the rank ordered population). The X-axis shows the average imaging value for the region analyzed. As can be seen in FIGS. 19-30, those individuals who fall generally in the middle of the population distribution (e.g., between the 25^(th) and 75^(th) percentile; the 33^(rd) and 67^(th) percentile; or the 40^(th) and 60^(th) percentile) exhibit color imaging values that can be used to distinguish the different types of periorbital dyschromia from one another.

FIGS. 19-30 are side-by-side comparisons of graphical plots representing imaging values corresponding to Type I, Type II and Type III test subjects. FIG. 19 shows a side-by-side comparison of Zone 1 R values. FIG. 20 shows a side-by-side comparison of the Zone 1 B values. FIG. 21 shows a side-by-side comparison of the Zone 1 G values. FIG. 22 shows a side-by-side comparison of the Zone 1 B/G ratios. FIG. 23 shows a side-by-side comparison of the Zone 2 R values. FIG. 24 shows a side-by-side comparison of the Zone 2 G values. FIG. 25 shows a side-by-side comparison of the Zone 2 B values. FIG. 26 shows a side-by-side comparison of the Zone 2 B/G ratios. FIG. 27 shows a side-by-side comparison of the R values from Zone 4. FIG. 28 shows a side-by-side comparison of the B values from Zone 4. FIG. 29 shows a side-by-side comparison of the plots of the G values from Zone 4. FIG. 30 shows a side-by-side comparison of the plots of the B/G ratios from Zone 4.

Because skin is complex, there can be many different colors in a particular region of interest. Different people may appear to have some or all of the same colors present in particular area of skin, but the colors may be present at different percentages. And it is the distribution and/or preponderance of the different colors in the region of interest that drives how periorbital dyschromia is perceived. Thus, the images described in this Example were also analyzed on a pixel by pixel basis to determine if differences between different types of periorbital dyschromia could be observed. The RGB values for each pixel were converted to LabCh values as described in the imaging method above, and the L, C and h values were normalized to the cheek basal skin tone. The DL, Dc and Dh values of each pixel are classified as follows:

DL>−5: the pixel is not meaningfully darker than the nearby cheek skin (or it could even be lighter), these pixels were classified as No Dyschromia

DL<−5 and DC>2: the pixel is both darker and more chromatic than the nearby skin. These pixels are classified as Type I.

DL<−5 and DC<2: the pixel is darker, but without a meaningful increase in chroma. These pixels are classified as Type II periorbital dyschromia.

After each pixel was classified, the distribution of the pixels in the masked region was determined by counting the number of pixels classified as each type of periorbital dyschromia (i.e., No Dyschromia, Type I and Type II) and dividing by the total number pixels in each mask to obtain percentages, (e.g., % Type I=# pixels classified as Type I divided by Total # pixels) Table 1 illustrates how the different types of periorbital dyschromia may be classified based on the observations from the pixel by pixel analysis of the images.

TABLE 4 Subject Pixel Distribution Classified as: >36.4% classified as Type I Type I <9.6% classified as Type I; AND Type II >21.2% classified as Type II >9.6% but less than 36.4% Type III classified as Type I; AND >20% classified as Type II. <9.6% classified as Type I AND No <21.2% classified as Type II. Dyschromia

Example 3

This example further illustrates how imaging values can be used to distinguish Type I, Type II and Type III periorbital dyschromia. Using the methods described herein, images were obtained from test subjects identified as having Type I, Type II, or Type III periorbital dyschromia, and the color corrected imaging values for each of Mask A, Mask B and Mask C were determined for each image. Table 5 shows the resulting values.

TABLE 5 Type I Imaging Value Mask A Mask B Mask C R 153 167 161 G 108 123 116 B 95 101 98 B/G 0.87 0.82 0.84 L 50 55 53 a* 16 14 15 b* 15 19 17 Hue (h) 43 53 49 Chroma (C) 22 23 23

TABLE 6 Type II Imaging Value Mask A Mask B Mask C R 161 176 169 G 119 135 128 B 108 116 112 B/G 0.91 0.86 0.88 L 54 60 57 a* 15 13 14 b* 12 17 15 Hue (h) 39 52 46 Chroma (C) 20 21 20

TABLE 7 Type III Imaging Value Mask A Mask B Mask C R 157 176 167 G 113 133 124 B 101 111 107 B/G 0.89 0.84 0.86 L 52 59 56 a* 16 13 14 b* 14 18 16 Hue (h) 41 54 48 Chroma (C) 21 23 22

This example illustrates the histological differences between No Dyschromia, Type I, Type II and Type III periorbital dyschromia. Biopsy samples were collected from upper eyelid, lower eyelid and the area of the face near the temple of fifty-five test subjects. The biopsy samples were collected, stored, sectioned and stained with H&E stain according to the Biopsy method above. An analysis of the sectioned and stained samples revealed the presence of deposits disposed around the nuclei of cells clustered together in the dermal region of the tissue sample. The deposits appeared brown in a color micrograph, and it is believed that the deposits are melanin. The samples were stained with Fontana-Masson stain and turned black (i.e., “Fontana-Masson positive deposits”), which indicates that the deposits may be melanin.

Thirty biopsy samples representative of the four different classifications were selected at random for further analysis (6 non-dyschromia samples, 9 Type I sample, 9 Type II samples, and 6 Type III samples). The thirty samples were analyzed at 20× magnification under an Olympus® Model BX51TRF brand light microscope. For the analysis, a minimum of two tissue sections and 8 fields of view (“FOV”), which is defined as the area of tissue that can be observed under 20× magnification, are analyzed per subject. The analysis revealed that Type I and Type III periorbital dyschromia exhibited higher amounts of the Fontana-Masson positive deposits on the dermis than Type II and those subjects identified as not having severe periorbital dyschromia. The average number of Fontana-Masson positive deposits per field of view is provided in Table 2 below. FIG. 12 graphically depicts the results illustrated in Table 2.

TABLE 8 No Dyschromia Type I Type II Type III Subject 1 2.88 15.00 0.87 13.6 Subject 2 4.67 11.50 0.10 4.19 Subject 3 1.69 22.45 0.00 2.35 Subject 4 1.94 12.96 4.00 3.81 Subject 5 2.58 11.50 0.38 6.33 Subject 6 2.33 6.95 0.56 14.25 Subject 7 10.58 0.07 Subject 8 12.33 0.00 Subject 9 21.78 0.50 Average 2.68 13.89 0.75 7.42 STD DEV 0.43 1.49 0.45 2.12

As can be seen in Table 8 and FIG. 12, the Fontana-Masson positive deposits distinguish the different types of periorbital dyschromia from each other and from the control. The chart in FIG. 12 illustrates this distinction based on the difference in the average number of Fontana-Masson positive deposits observed per FOV under 20× magnification.

Example 4

This example illustrates the differences in gene expression between No Dyschromia, Type I, Type II and Type III periorbital dyschromia. In this example, gene expression profiles and phenotype themes were determined from biopsy samples of female test subjects aged 18 to 45 who were classified as exhibiting Type I, Type II or Type III periorbital dyschromia by an expert grader. Thirteen subjects were classified as Type I periorbital dyschromia, fourteen subjects were classified as Type II periorbital dyschromia, seven subjects were classified as Type III periorbital dyschromia, and 8 test subjects were classified as No Dyschromia. The biopsy samples were collected, stored and sectioned according to the Biopsy method described above. The epidermis and dermis layers of the section biopsy samples were separated with a PALM Microbeam IV™ brand Laser-capture Micro dissection (“LCM”) system (available from Carl Zeiss MicroImaging GmbH, Germany) in accordance with the manufacturer's instructions.

After separating the epidermal and dermal layers of the sectioned biopsy samples, each layer was subjected to RNA extraction. In this example, RNA was extracted from each of the epidermis and dermis layers by utilizing the Arcturs picopure kit according to the manufacturer's instructions. RNA quantification and quality assurance was performed using Agilence 2100 bioanalyzer. Of course it is to be appreciated that any suitable means of extracting RNA from a tissue sample known in the art may be used. The extracted RNA was then run on a GeneTitan U129 brand microarray to identify how certain genes were expressed in each sample. A statistical analysis of the microarray data was performed to derive the gene expression signatures. A general description of the statistical analysis is provided below. It is to be appreciated that while the statistical analysis below provides a suitable means of generating a gene signature from the microarray data, other statistical methods known in the art may also be suitable for use herein.

Filtering based on Present Calls

Filtering creates a list of potential genes for inclusion in the gene expression signature. For example, as a first filter step, at least 50% of the samples in one treatment group must have a “Present call” for each probe set. Present calls are derived from processing the raw microarray data and provide evidence that the gene transcript complementary to a probe set is actually expressed in the biological sample. The probes that are absent from all samples are likely to be just noisy measurements. It is important to filter out probe sets that do not contribute meaningful data to the signature. For all gene expression signatures in this example, the data was filtered for probe sets with at least 10% Present calls provided by the Affymetrix MAS 5 software.

Filtering According to a Statistical Measure

As a second filtering step, it may be desirable to use a suitable statistical measure such as, for example, p-values from a t-test, ANOVA, correlation coefficient, or other suitable model-based analysis. Limiting the gene signature list to genes that meet some reasonable cutoff (e.g., p ≤0.05, 0.01, 0.001, or even ≤0.0001 or less) for statistical significance compared to an appropriate control is important to allow selection of genes that are characteristic of the biological state of interest. This is preferable to using a fold change value, which does not take into account the noise around the measurements. For example, p-values may be chosen as the statistical measure and a cutoff value of p≤0.05 may be chosen. The t-statistic was used in this example to select the probe sets in the signatures because it provides an indication of the directionality of the gene expression changes (i.e. up- or down-regulated) as well as statistical significance.

Sorting the Probe Sets

All the probe sets are sorted into sets of up-regulated and down-regulated sets using the statistical measure. In this example, a t-test was used to compute p-values, the values (positive and negative) of the t-statistic are used to sort the list since p-values are always positive. The sorted t-statistics will place the sets with the most significant p-values at the top and bottom of the list with the less-significant p-values being placed towards the middle.

Creation of the Gene Expression Signature

Using the filtered and sorted list created, a suitable number of probe sets from the top and bottom are selected to create a gene expression signature that preferably has approximately the same number of sets chosen from the top as chosen from the bottom. For example, the gene expression signature created may have at least 10, 50, 100, 200, or 300 and/or less than 800, 600, or about 400 genes corresponding to a probe set on the chip. The number of probe sets approximately corresponds to the number of genes, but a single gene may be represented by more than one probe set. It is understood that the phrase “number of genes” as used herein, corresponds generally with the phrase “number of probe sets.”

U.S. Publication No. 2012/0283112 titled “Systems and Methods For Identifying Cosmetic Agents For Skin Care Compositions” filed by Binder, et al., on Feb. 22, 2012 and U.S. Publication No. 2013/0261007 titled “Systems, Models and Methods for Identifying and Evaluating Skin-Active Agents Effective for Treating Conditions and Disorders of Skin Pigmentation,” filed by Hakozaki, et al., on Mar. 27, 2012 disclose suitable nonlimiting examples of methods of generating a gene expression profile.

Tables 9 through 20 below show gene expression signatures associated with Type I, Type II or Type III periorbital dyschromia, as compared to the gene expression signature for No Dyschromia. The gene expression data was obtained by extracting RNA from lower eyelid biopsy samples obtained according to the biopsy method. Only genes that showed up-regulation or down-regulation with a p-value of 0.05 or less and fold change of 1.1 or more are shown. Table 9 shows the top 100 up-regulated genes expressed in the epidermis of subjects identified as having Type I periorbital dyschromia. Table 10 shows the top 100 down-regulated genes expressed in the epidermis of subjects identified as having Type I periorbital dyschromia. Table 11 shows the top 100 up-regulated genes expressed in the dermis of subjects identified as having Type I periorbital dyschromia. Table 12 shows the top 100 down-regulated genes expressed in the dermis of subjects identified as having Type I periorbital dyschromia. Table 13 shows the top 100 up-regulated genes expressed in the epidermis of subjects identified as having Type II periorbital dyschromia. Table 14 shows the top 100 down-regulated genes expressed in the dermis of subjects identified as having Type II periorbital dyschromia. Table 15 shows the top 71 up-regulated genes expressed in the dermis of subjects identified as having Type II periorbital dyschromia. Table 16 shows the top 100 down-regulated genes expressed in the dermis of subjects identified as having Type II periorbital dyschromia. Table 17 shows the top 100 up-regulated genes expressed in the epidermis of subjects identified as having Type III periorbital dyschromia. Table 18 shows the top 100 down-regulated genes expressed in the epidermis of subjects identified as having Type III periorbital dyschromia. Table 19 shows the top 100 up-regulated genes expressed in the dermis of subjects identified as having Type III periorbital dyschromia. Table 20 show the top 100 down-regulated genes expressed in the dermis of subjects identified as having Type III periorbital dyschromia.

TABLE 9 Type I Epidermis; Up-regulated GeneTitan_ID Gene Title Public ID p 11719428_a_at BTBD7 BTB (POZ) domain containing 7 AI580162 0.000259 11724297_a_at BMP7 bone morphogenetic protein 7 NM_001719.2 0.000396 11715192_s_at C7orf46 chromosome 7 open reading frame 46 g188219621 0.000446 11724759_s_at CALM1 calmodulin 1 (phosphorylase kinase, NM_006888.3 0.000486 delta) 11735610_a_at RPS6KA5 ribosomal protein S6 kinase, 90 kDa, AF074393.1 0.000746 polypeptide 5 11758022_s_at TNNI2 troponin I type 2 (skeletal, fast) AA728828 0.00151 11721260_a_at WDR47 WD repeat domain 47 NM_001142550.1 0.001528 11727979_a_at MAN2B2 mannosidase, alpha, class 2B, BC094773.1 0.002024 member 2 11753967_a_at SLC6A2 solute carrier family 6 AK301811.1 0.002393 (neurotransmitter transporter, noradrenalin), member 2 11722090_a_at EFNA4 ephrin-A4 NM_005227.2 0.002687 11758092_s_at EFNA5 ephrin-A5 BE464799 0.002776 11717146_at PTPN1 protein tyrosine phosphatase, non- NM_002827.2 0.002876 receptor type 1 11716283_at PAPD7 PAP associated domain containing 7 NM_006999.3 0.003497 11746140_a_at ARHGEF26 Rho guanine nucleotide exchange AF415176.1 0.003751 factor (GEF) 26 11724038_a_at PTGS2 prostaglandin-endoperoxide BC013734.1 0.003867 synthase 2 (prostaglandin G/H synthase and cyclooxygenase) 11723156_a_at LSP1 lymphocyte-specific protein 1 NM_001013254.1 0.004011 11729840_s_at ZCCHC2 zinc finger, CCHC domain containing 2 NM_017742.4 0.004086 11754302_a_at ATP2A2 ATPase, Ca++ transporting, cardiac BM676899 0.004156 muscle, slow twitch 2 11754447_a_at RPS6KA5 ribosomal protein S6 kinase, 90 kDa, BM968829 0.00436 polypeptide 5 11739724_a_at ATP2B2 ATPase, Ca++ transporting, plasma AL138283 0.004438 membrane 2 11723075_a_at BCL9L B-cell CLL/lymphoma 9-like AY296059.1 0.00493 11717385_a_at MT1G metallothionein 1G NM_005950.1 0.005447 11758557_s_at ZFP36L1 zinc finger protein 36, C3H type-like 1 AI758505 0.005544 11724628_a_at MAT1A methionine adenosyltransferase I, NM_000429.2 0.005576 alpha 11744955_a_at ANXA1 annexin A1 AK296808.1 0.005629 11718966_at NUFIP2 nuclear fragile X mental retardation BU533767 0.006124 protein interacting protein 2 11744953_a_at ANXA1 annexin A1 BC034157.1 0.007553 11717387_x_at MT1G metallothionein 1G NM_005950.1 0.007682 11727642_a_at TRERF1 transcriptional regulating factor 1 AF111801.1 0.0078 11745806_a_at AMMECR1L AMME chromosomal region gene 1-like AK095871.1 0.008163 11717514_a_at ANXA1 annexin A1 NM_000700.1 0.008725 11744954_x_at ANXA1 annexin A1 BC034157.1 0.009223 11743864_a_at ATP2B1 ATPase, Ca++ transporting, plasma S49852.1 0.009249 membrane 1 11717927_at FOXK2 forkhead box K2 NM_004514.3 0.009268 11717386_s_at MT1G metallothionein 1G NM_005950.1 0.009421 11730080_x_at IL28RA interleukin 28 receptor, alpha NM_170743.2 0.009429 (interferon, lambda receptor) 11763297_x_at CCDC76 coiled-coil domain containing 76 BQ614335 0.010042 11726119_at RPGR retinitis pigmentosa GTPase regulator AK291832.1 0.01012 11739805_a_at RASAL2 RAS protein activator like 2 AK075169.1 0.010243 11715190_s_at C7orf46 chromosome 7 open reading frame 46 g188219617 0.010448 11726351_at EFNA5 ephrin-A5 CB240929 0.010459 11738035_s_at RTN4 reticulon 4 AK302741.1 0.010618 11724104_s_at SGK3 serum/glucocorticoid regulated NM_001033578.1 0.010735 kinase family, member 3 11729396_a_at NEK1 NIMA (never in mitosis gene a)- Z25431.1 0.010781 related kinase 1 11723592_at LRRC8C leucine rich repeat containing 8 NM_032270.4 0.011015 family, member C 11757982_s_at KIF21A kinesin family member 21A N39407 0.011191 11731899_s_at PPAT phosphoribosyl pyrophosphate D13757.1 0.01137 amidotransferase 11726633_s_at TRIM8 tripartite motif-containing 8 BC021925.1 0.011932 11725675_a_at RORA RAR-related orphan receptor A AA034012 0.012081 11745021_a_at MYC v-myc myelocytomatosis viral K02276.1 0.012276 oncogene homolog (avian) 11758246_s_at ARL4D ADP-ribosylation factor-like 4D BM719529 0.012438 11726634_a_at MYST3 MYST histone acetyltransferase NM_001099412.1 0.012468 (monocytic leukemia) 3 11735421_a_at NKD2 naked cuticle homolog 2 AF358137.1 0.013343 (Drosophila) 11723821_a_at SMURF2 SMAD specific E3 ubiquitin protein AY014180.1 0.013348 ligase 2 11733165_s_at YIPF5 Yip1 domain family, member 5 NM_001024947.2 0.013814 11723169_s_at FOXN2 forkhead box N2 NM_002158.3 0.014398 11728958_x_at E2F8 E2F transcription factor 8 NM_024680.2 0.014717 11762525_s_at AF339086.1 0.015079 11731645_a_at CAMKK2 calcium/calmodulin-dependent BC026060.2 0.016269 protein kinase kinase 2, beta 11730893_a_at UBA6 ubiquitin-like modifier activating EF623993.1 0.016784 enzyme 6 11730360_at CCDC126 coiled-coil domain containing 126 NM_138771.3 0.016798 11736426_s_at CLIP4 CAP-GLY domain containing linker BM994685 0.017149 protein family, member 4 11725820_s_at PAQR3 progestin and adipoQ receptor NM_001040202.1 0.01721 family member III 11743411_a_at RBM25 RNA binding motif protein 25 BG251218 0.017641 11718873_a_at ATP2A2 ATPase, Ca++ transporting, cardiac NM_170665.3 0.018009 muscle, slow twitch 2 11739797_a_at RFX2 regulatory factor X, 2 (influences NM_134433.2 0.018119 HLA class II expression) 11739408_at MLL3 myeloid/lymphoid or mixed-lineage DN917896 0.018481 leukemia 3 11734164_a_at ARHGEF26 Rho guanine nucleotide exchange NM_015595.3 0.018897 factor (GEF) 26 11721039_a_at SOLH small optic lobes homolog NM_005632.2 0.019352 (Drosophila) 11721053_s_at KLHDC5 kelch domain containing 5 NM_020782.1 0.01948 11753735_x_at TMSB4X thymosin beta 4, X-linked BC101792.1 0.019698 11748149_a_at FNBP1 formin binding protein 1 AK293743.1 0.019714 11728706_x_at EMP2 epithelial membrane protein 2 DB374012 0.020127 11717989_a_at SUN1 Sad1 and UNC84 domain containing 1 NM_001130965.1 0.020293 11722448_at KCNMB4 potassium large conductance AF160967.1 0.020548 calcium-activated channel, subfamily M, beta member 4 11727512_at UBN2 ubinuclein 2 CA775887 0.020981 11759962_at TPRKB TP53RK binding protein AY643713.1 0.021007 11728603_a_at CLDN23 claudin 23 NM_194284.2 0.0211 11743497_at BMP2 bone morphogenetic protein 2 BX101090 0.021788 11718874_s_at ATP2A2 ATPase, Ca++ transporting, cardiac NM_170665.3 0.0218 muscle, slow twitch 2 11721216_s_at TMEM106B transmembrane protein 106B AA789109 0.021968 11718006_a_at MYLIP myosin regulatory light chain BC002860.2 0.02198 interacting protein 11719104_s_at CPNE3 copine III BC066597.1 0.022194 11723130_a_at ARHGEF7 Rho guanine nucleotide exchange NM_003899.3 0.022803 factor (GEF) 7 11757700_a_at NDFIP2 Nedd4 family interacting protein 2 AA521251 0.022979 11718081_a_at ATP2B4 ATPase, Ca++ transporting, plasma NM_001001396.1 0.023666 membrane 4 11748034_a_at CMAH cytidine monophosphate-N- D86324.1 0.02381 acetylneuraminic acid hydroxylase (CMP-N-acetylneuraminate monooxygenase) pseudogene 11740148_x_at ZNF429 zinc finger protein 429 NM_001001415.2 0.023983 11716408_a_at MET met proto-oncogene (hepatocyte NM_001127500.1 0.024171 growth factor receptor) 11720082_at CBX6 chromobox homolog 6 NM_014292.3 0.024178 11733076_x_at PPP1R12A protein phosphatase 1, regulatory AK314193.1 0.024224 (inhibitor) subunit 12A 11743280_a_at WNK1 WNK lysine deficient protein kinase 1 BC013629.2 0.024246 11719028_a_at PSD3 pleckstrin and Sec7 domain DB314358 0.024644 containing 3 11757869_s_at AKAP13 A kinase (PRKA) anchor protein 13 BE504033 0.024753 11722290_a_at ZBTB43 zinc finger and BTB domain AI745225 0.024875 containing 43 11743865_s_at ATP2B1 ATPase, Ca++ transporting, plasma AI337321 0.025057 membrane 1 11734994_s_at SKI v-ski sarcoma viral oncogene NM_003036.3 0.025378 homolog (avian) 11731857_x_at MT1H metallothionein 1H NM_005951.2 0.026268 11758247_x_at ARL4D ADP-ribosylation factor-like 4D BM719529 0.026757 11731787_x_at ERC1 ELKS/RAB6-interacting/CAST family NM_178037.1 0.026811 member 1

TABLE 10 Type I Epidermis; Down-regulated GeneTitan_ID Gene Title Public ID p 11763233_x_at TRAC T cell receptor alpha constant EU427374.1 0.000086 11750712_a_at SEMA4A sema domain, immunoglobulin AK296693.1 0.000205 domain (Ig), transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 4A 11743350_a_at C15orf48 chromosome 15 open reading CA309087 0.000277 frame 48 11723817_at ARHGAP29 Rho GTPase activating protein 29 BU620659 0.000376 11723854_at SAMD9 sterile alpha motif domain NM_017654.2 0.000398 containing 9 11754862_a_at RARRES2 retinoic acid receptor AK092804.1 0.000534 responder (tazarotene induced) 2 11729515_a_at SLC26A9 solute carrier family 26, NM_052934.3 0.001204 member 9 11748896_s_at CCRL1 chemokine (C-C motif) AK304461.1 0.001209 receptor-like 1 11761960_x_at TRAV20 T cell receptor alpha variable 20 AY532913.1 0.001367 11741285_a_at CCRL1 chemokine (C-C motif) BC069438.1 0.00144 receptor-like 1 11754482_a_at PSMB1 proteasome (prosome, BQ006450 0.001693 macropain) subunit, beta type, 1 11754184_a_at ALDH1A3 aldehyde dehydrogenase 1 BX538027.1 0.001883 family, member A3 11726829_at TYW1B tRNA-yW synthesizing protein 1 NM_001145440.1 0.001956 homolog B (S. cerevisiae) 11725368_at LRG1 leucine-rich alpha-2- NM_052972.2 0.001997 glycoprotein 1 11723561_x_at C11orf75 chromosome 11 open reading NM_020179.2 0.002048 frame 75 11734507_s_at MECOM MDS1 and EVI1 complex locus AK292865.1 0.002142 11716805_s_at PLEKHA2 pleckstrin homology domain NM_021623.1 0.002256 containing, family A (phosphoinositide binding specific) member 2 11754057_x_at CRABP2 cellular retinoic acid binding BT019827.1 0.00239 protein 2 11737108_a_at CCRL1 chemokine (C-C motif) NM_178445.1 0.002438 receptor-like 1 11720080_at NTRK2 neurotrophic tyrosine kinase, NM_001007097.1 0.002467 receptor, type 2 11715767_s_at ACAA2 acetyl-CoA acyltransferase 2 NM_006111.2 0.002477 11715670_a_at IFITM1 interferon induced NM_003641.3 0.002619 transmembrane protein 1 (9-27) 11735937_a_at CD48 CD48 molecule NM_001778.2 0.002662 11746503_a_at EHF ets homologous factor AF203977.1 0.00321 11736806_at GABRA4 gamma-aminobutyric acid NM_000809.2 0.003627 (GABA) A receptor, alpha 4 11720157_at GDA guanine deaminase AK295716.1 0.003854 11720132_a_at SPIRE1 spire homolog 1 (Drosophila) BC125206.1 0.003917 11725832_s_at OTUB2 OTU domain, ubiquitin NM_023112.3 0.004105 aldehyde binding 2 11717763_a_at MGLL monoglyceride lipase BC006230.2 0.004157 11756683_a_at CD1E CD1e molecule AK311643.1 0.004158 11728008_x_at FUT3 fucosyltransferase 3 NM_000149.3 0.004308 (galactoside 3(4)-L- fucosyltransferase, Lewis blood group) 11750244_a_at MGLL monoglyceride lipase AK304844.1 0.004332 11724346_a_at IFIH1 interferon induced with NM_022168.2 0.004365 helicase C domain 1 11750245_x_at MGLL monoglyceride lipase AK304844.1 0.004459 11758377_s_at TLR1 toll-like receptor 1 BU623316 0.004771 11740897_a_at TREX2 three prime repair exonuclease 2 NM_080701.3 0.004847 11718077_s_at MAPKAPK3 mitogen-activated protein NM_004635.3 0.004908 kinase-activated protein kinase 3 11763697_s_at SNHG9 small nucleolar RNA host gene AW958849 0.004956 9 (non-protein coding) 11719591_s_at GLTP glycolipid transfer protein NM_016433.3 0.005188 11715671_x_at IFITM1 interferon induced NM_003641.3 0.005216 transmembrane protein 1 (9- 27) 11725641_at EFHD2 EF-hand domain family, CB240768 0.005381 member D2 11727633_at SLC16A10 solute carrier family 16, BC066985.1 0.005429 member 10 (aromatic amino acid transporter) 11717764_x_at MGLL monoglyceride lipase BC006230.2 0.005967 11750324_a_at GAS7 growth arrest-specific 7 AK293755.1 0.006052 11735833_a_at KIAA1199 KIAA1199 NM_018689.1 0.006555 11757367_s_at HSPA7 heat shock 70 kDa protein 7 BM677874 0.006575 (HSP70B) 11741263_s_at LTBP1 latent transforming growth M34057.1 0.006584 factor beta binding protein 1 11723235_a_at IFI44L interferon-induced protein 44-like AB000115.1 0.006657 11729692_a_at SERPINB3 serpin peptidase inhibitor, EU852041.1 0.006806 clade B (ovalbumin), member 3 11727385_a_at PCCA propionyl CoA carboxylase, NM_000282.2 0.006811 alpha polypeptide 11729742_x_at IFI27L2 interferon, alpha-inducible NM_032036.2 0.006823 protein 27-like 2 11731973_at SCNN1G sodium channel, nonvoltage- NM_001039.3 0.00716 gated 1, gamma 11743404_at ZMAT2 zinc finger, matrin-type 2 BM450158 0.007256 11736058_s_at C10orf32 chromosome 10 open reading NM_001136200.1 0.007371 frame 32 11715239_x_at IFITM3 interferon induced g148612841 0.007374 transmembrane protein 3 (1-8U) 11726154_at CA6 carbonic anhydrase VI NM_001215.2 0.007409 11718850_a_at SRPK1 SRSF protein kinase 1 AK299591.1 0.007419 11729693_at SERPINB3 serpin peptidase inhibitor, EU852041.1 0.007514 clade B (ovalbumin), member 3 11755950_a_at coiled-coil domain containing 71 AK098658.1 0.007621 11740349_at RNASE7 ribonuclease, RNase A family, 7 BC112334.1 0.007692 11757300_s_at ELOVL5 ELOVL family member 5, AL576414 0.007702 elongation of long chain fatty acids (FEN1/Elo2, SUR4/Elo3- like, yeast) 11725310_at CRISP3 cysteine-rich secretory protein 3 NM_006061.1 0.007915 11723490_at GCLM glutamate-cysteine ligase, BC041809.1 0.008288 11757595_x_at CRABP2 cellular retinoic acid binding BU631189 0.008352 protein 2 11752101_s_at EIF2S1 eukaryotic translation initiation BC002513.2 0.008373 factor 2, subunit 1 alpha, 35 kDa 11737496_a_at CD200R1 CD200 receptor 1 NM_170780.2 0.008378 11749745_a_at SRP68 signal recognition particle AK301100.1 0.008462 68 kDa 11718142_a_at TTC27 tetratripeptide repeat NM_017735.3 0.00876 domain 27 11737432_a_at PAPL iron/zinc purple acid BC136722.1 0.008796 phosphatase-like-protein 11753762_x_at KLK6 kallikrein-related peptidase 6 AY457039.1 0.008902 11725875_at WDR66 WD repeat domain 66 NM_144668.4 0.008916 11729694_s_at SERPINB4 serpin peptidase inhibitor, EU852041.1 0.009199 clade B (ovalbumin), member 4 11763184_at IDE insulin-degrading enzyme BQ006777 0.009364 11737743_a_at ARSF arylsulfatase F NM_004042.3 0.009395 11724785_x_at MRPS18C mitochondrial ribosomal BC005186.1 0.009553 protein S18C 11723899_a_at DHRS9 dehydrgenase/reductase (SDR NM_005771.4 0.009689 family) member 9 11743805_s_at MRPL42 mitochondrial ribosomal DB379276 0.009847 protein L42 11758083_s_at HPGD hydroxyprostaglandin AI743714 0.009851 dehydrogenase 15-(NAD) 11727995_a_at SPINKS serine peptidase inhibitor DQ149928.1 0.010277 Kazal type 5 11727208_x_at DDHD1 DDHD domain containing 1 NM_030637.1 0.010297 11737431_x_at PAPL iron/zinc purple acid NM_001004318.2 0.010375 phosphatase-like protein 11720510_a_at APOBEC3G apolopoprotein B mRNA editing NM_021822.2 0.010471 enzyme, catalytic polypeptide- like 3G 11719503_a_at DHX36 DEAH (Asp-Glu-Ala-His) box NM_020865.2 0.010702 polypeptide 36 11753152_x_at CFLAR CASP8 and FADD-like apoptosis AK297387.1 0.010705 regulator 11726289_at GRAMD3 GRAM domain containing 3 BC008590.1 0.010722 11722661_at NFE2L3 nuclear factor (erythroid- NM_004289.6 0.010893 derived 2)-like 3 11716743_s_at TJP2 tight junction protein 2 (zona NM_004817.2 0.011228 occludens 2) 11726894_a_at IRAK3 interleukin-1 receptor- BG929347 0.011364 associated kinase 3 11753202_s_at SERPINB4 serpin peptidase inhibitor, AB046400.1 0.011604 clade B (ovalbumin), member 4 11739292_at EHF ets homologous factor NM_012153.3 0.012013 11748253_a_at SLC5A1 solute carrier family 5 AK297665.1 0.01209 (sodium/glucose cotransporter), member 1 11723953_a_at CLINT1 clathrin interactor 1 NM_014666.2 0.01254 11736117_a_at ZFAND5 zinc finger, AN1-type domain 5 AF062346.1 0.012593 11724795_at ZG16B zymogen granule protein 16 NM_145252.2 0.013029 homolog B (rat) 11717765_a_at MGLL monoglyceride lipase NM_007283.5 0.01317 11744194_a_at ABCC3 ATP-binding cassette, sub- CB055248 0.013406 family C (CFTR/MRP), member 3 11746088_a_at IFI44 interferon-induced protein 44 DB350079 0.013466 11733533_at CYP4F22 cytochrome P450, family 4, NM_173483.3 0.013708 subfamily F, polypeptide 22 11746581_a_at PCCA propionyl CoA carboxylase, AK298318.1 0.013745 alpha polypeptide 11717981_a_at ACP5 acid phosphatase 5, tartrate NM_001611.3 0.014935 resistant

TABLE 11 Type I Dermis; Up-regulated GeneTitan_ID Gene Title Public ID p 11733167_at LRRN4CL LRRN4 C-terminal like BC053902.1 0.000112 11720616_a_at DNM1 dynamin 1 NM_001005336.1 0.000113 11758194_s_at DPP4 dipeptidyl-peptidase 4 AI768728 0.000125 11759481_at COPB1 Coatomer protein complex, AU143964 0.000148 subunit beta 1 11718627_at TRAK1 trafficking protein, kinesin CA415544 0.000173 binding 1 11739544_a_at C19orf12 chromosome 19 open reading BX328123 0.000234 frame 12 11743191_a_at NTM neurotrimin AI343272 0.000278 11731649_x_at NTM neurotrimin AY358331.1 0.000343 11723111_a_at EMILIN2 elastin microfibril interfacer 2 NM_032048.2 0.000384 11719422_s_at ABCC1 ATP-binding cassette, sub- NM_004996.3 0.000385 family C (CFTR/MRP), member 1 11739527_a_at SECTM1 secreted and transmembrane 1 CR614987.1 0.000402 11758810_at COL14A1 collagen, type XIV, alpha 1 NM_021110.1 0.000451 11724441_x_at PTGIS prostaglandin I2 NM_000961.3 0.000458 (prostacyclin) synthase 11747944_a_at PPFIA2 protein tyrosine phosphatase, AK296380.1 0.000483 receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 2 11728451_a_at PCOLCE2 procollagen C-endopeptidase NM_013363.2 0.000493 enhancer 2 11736086_a_at HHIP hedgehog interacting protein NM_022475.1 0.000511 11758252_s_at HSD3B7 hydroxy-delta-5-steroid CB115219 0.000562 dehydrogenase, 3 beta- and steroid delta-isomerase 7 11756706_a_at DPP4 dipeptidyl-peptidase 4 AK314798.1 0.000563 11720690_a_at C2orf18 chromosome 2 open reading NM_017877.3 0.000604 frame 18 11724619_at RSPO3 R-spondin 3 homolog NM_032784.3 0.000676 (Xenopus laevis) 11743447_a_at BICD2 bicaudal D homolog 2 AW409827 0.000721 (Drosophila) 11725773_a_at TBC1D24 TBC1 domain family, member 24 NM_020705.1 0.000726 11715704_x_at ITGA5 integrin, alpha 5 (fibronectin NM_002205.2 0.000766 receptor, alpha polypeptide) 11756007_a_at HHIP hedgehog interacting protein AK024645.1 0.000889 11741377_a_at MMP2 matrix metallopeptidase 2 NM_001127891.1 0.000928 (gelatinase A, 72 kDa gelatinase, 72 kDa type IV collagenase) 11734550_x_at TGFBI transforming growth factor, NM_000358.2 0.000963 beta-induced, 68 kDa 11730405_at MEX3B mex-3 homolog B (C. elegans) BC111545.1 0.000968 11731648_a_at NTM neurotrimin AY358331.1 0.000985 11740103_a_at MAFG v-maf musculoaponeurotic BX427058 0.001077 fibrosarcoma oncogene homolog G (avian) 11727783_s_at TPM4 tropomyosin 4 NM_003290.2 0.001163 11718269_x_at ANGPTL2 angiopoietin-like 2 AY358274.1 0.001231 11725897_at TUBB1 tubulin, beta 1 BC033679.1 0.001234 11717803_a_at NTN4 netrin 4 NM_021229.3 0.001243 11754476_x_at DNM1 dynamin 1 BQ183716 0.00125 11746893_a_at MPP1 membrane protein, AK304538.1 0.001254 palmitoylated 1, 55 kDa 11753088_a_at MCTP1 multiple C2 domains, AK297325.1 0.001305 transmembrane 1 11717568_s_at NQ01 NAD(P)H dehydrogenase, NM_000903.2 0.001408 quinone 1 11720051_at SPOCK1 sparc/osteonectin, cwcv and NM_004598.3 0.001423 kazal-like domains proteoglycan (testican) 1 11757921_s_at COL14A1 collagen, type XIV, alpha 1 AI248460 0.001424 11725923_s_at ASAP2 ArfGAP with SH3 domain, NM_001135191.1 0.001457 ankyrin repeat and PH domain 2 11723225_a_at CLDN11 claudin 11 BC013577.1 0.001495 11754368_a_at FBN1 fibrillin 1 AB208840.1 0.001626 11758062_s_at STK32B serine/threonine kinase 32B AI401203 0.001735 11740358_a_at LILRB5 leukocyte immunoglobulin- NM_001081443.1 0.001775 like receptor, subfamily B (with TM and ITIM domains), member 5 11746200_s_at EHD2 EH-domain containing 2 AK097126.1 0.001844 11747945_x_at PPFIA2 protein tyrosine phosphatase, AK296380.1 0.002036 receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 2 11720811_a_at PAMR1 peptidase domain containing NM_015430.2 0.002139 associated with muscle regeneration 1 11731716_at CCBP2 chemokine binding protein 2 NM_001296.4 0.002156 11754706_a_at HHIP hedgehog interacting protein AK098525.1 0.002166 11722940_a_at SRGAP2 SLIT-ROBO Rho GTPase NM_001042758.1 0.00222 activating protein 2 11724848_a_at DIXDC1 DIX domain containing 1 DB358954 0.002285 11718842_a_at C16orf62 chromosome 16 open reading BC058845.1 0.002303 frame 62 11734549_s_at TGFBI transforming growth factor, NM_000358.2 0.002382 beta-induced, 68 kDa 11734548_a_at TGFBI transforming growth factor, NM_000358.2 0.002425 beta-induced, 68 kDa 11731650_a_at NTM neurotrimin NM_001048209.1 0.002477 11738845_x_at NTM neurotrimin NM_001144059.1 0.002642 11726905_a_at ARHGAP20 Rho GTPase activating protein 20 NM_020809.2 0.002644 11724735_a_at PDPN podoplanin BC014668.1 0.002735 11755796_a_at ADAM9 ADAM metallopeptidase BC143924.1 0.002822 domain 9 11726017_a_at C17orf63 chromosome 17 open reading AU253346 0.002907 frame 63 11741286_a_at CCRL1 chemokine (C-C motif) AF110640.1 0.002908 receptor-like 1 11743910_at FAM69A family with sequence BQ015316 0.002927 similarity 69, member A 11756911_a_at C1QTNF3 C1q and tumor necrosis factor BX640995.1 0.002969 related protein 3 11746361_a_at C7orf58 chromosome 7 open reading BC030538.2 0.002971 frame 58 11715703_s_at ITGA5 integrin, alpha 5 (fibronectin NM_002205.2 0.002995 receptor, alpha polypeptide) 11735913_s_at TNXB tenascin XB BC125114.1 0.003064 11730236_s_at MYADM myeloid-associated AY358582.1 0.003084 differentiation marker 11718267_a_at ANGPTL2 angiopoietin-like 2 NM_012098.2 0.003114 11723217_x_at SFXN3 sideroflexin 3 NM_030971.3 0.003152 11720286_a_at TRAK1 trafficking protein, kinesin BC015922.1 0.003236 binding 1 11717133_a_at MAFG v-maf musculoaponeurotic BF340448 0.003272 fibrosarcoma oncogene homolog G (avian) 11717340_at PTGFRN prostaglandin F2 receptor NM_020440.2 0.003311 negative regulator 11729541_a_at CAMKK2 calcium/calmodulin- AB081337.1 0.003359 dependent protein kinase kinase 2, beta 11718268_a_at ANGPTL2 angiopoietin-like 2 AY358274.1 0.003373 11717413_a_at WIPI1 WD repeat domain, NM_017983.5 0.003406 phosphoinositide interacting 1 11716226_a_at LIMA1 LIM domain and actin binding 1 BC136763.1 0.003456 11740588_at BDKRB2 bradykinin receptor B2 NM_000623.3 0.003464 11741128_a_at CAPN2 calpain 2, (m/II) large subunit NM_001146068.1 0.003473 11717891_a_at ECM1 extracellular matrix protein 1 BC023505.2 0.003509 11730385_at GREM2 gremlin 2 BG150451 0.003541 11756245_s_at ANXA5 annexin A5 CR607543.1 0.003586 11721499_x_at CTSA cathepsin A NM_001127695.1 0.003681 11717757_s_at RALA v-ral simian leukemia viral AA548928 0.003773 oncogene homolog A (ras related) 11723075_a_at BCL9L B-cell CLL/lymphoma 9-like AY296059.1 0.003835 11748650_a_at ADAM33 ADAM metallopeptidase BC125113.1 0.003846 domain 33 11758676_s_at RHOQ ras homolog gene family, R23125 0.003853 member Q 11724260_a_at TRIO triple functional domain AF091395.1 0.004049 (PTPRF interacting) 11724541_s_at VWF von Willebrand factor NM_000552.3 0.004078 11716549_s_at ISLR immunoglobulin superfamily NM_005545.3 0.0042 containing leucine-rich repeat 11724228_at RBMS1 RNA binding motif, single BC080620.1 0.004204 stranded interacting protein 1 11752423_a_at F13A1 coagulation factor XIII, A1 AK304335.1 0.004293 polypeptide 11757340_s_at RHOQ ras homolog gene family, BM677515 0.004317 member Q 11750650_a_at PAMR1 peptidase domain containing AK297092.1 0.004367 associated with muscle regeneration 1 11735263_s_at SCN2A sodium channel, voltage- NM_001040142.1 0.004453 gated, type II, alpha subunit 11731682_at CD70 CD70 molecule NM_001252.3 0.004492 11737108_a_at CCRL1 chemokine (C-C motif) NM_178445.1 0.004501 receptor-like 1 11743251_s_at MMP2 matrix metallopeptidase 2 BX357054 0.004507 (gelatinase A, 72 kDa gelatinase, 72 kDa type IV collagenase) 11727782_a_at TPM4 tropomyosin 4 NM_003290.2 0.004515 11755955_a_at FAP fibroblast activation protein, AL832166.1 0.004528 alpha 11725868_at SSC5D scavenger receptor cysteine- NM_001144950.1 0.004768 rich glycoprotein

TABLE 12 Type I Dermis; Down-regulated GeneTitan_ID Gene Title Public ID p 11718273_a_at EIF3L eukaryotic translation NM_016091.2 0.000001 initiation factor 3, subunit L 11729152_a_at EIF3M eukaryotic translation NM_006360.3 0.000006 initiation factor 3, subunit M 11755203_x_at RPL21 ribosomal protein L21 BX647669.1 0.000006 11757356_x_at RPL30 ribosomal protein L30 BM855760 0.000008 11717236_x_at RPS7 ribosomal protein S7 NM_001011.3 0.000009 11745362_x_at RPS11 ribosomal protein S11 BC100025.1 0.000009 11757363_x_at RPS15A ribosomal protein S15a DB313157 0.000009 200062_PM_s_at RPL30 ribosomal protein L30 L05095.1 0.000009 11716092_x_at CKS1B CDC28 protein kinase NM_001826.2 0.00001 regulatory subunit 1B 11743094_at SPRR4 small proline-rich protein 4 BC069445.1 0.000011 11757421_x_at RPL31 ribosomal protein L31 CD687230 0.000011 11718274_s_at EIF3L eukaryotic translation NM_016091.2 0.000012 initiation factor 3, subunit L 11715376_a_at RPS11 ribosomal protein S11 NM_001015.3 0.000015 11757773_x_at NCRNA00275 non-protein coding RNA 275 BF185165 0.000015 11753659_x_at RPL30 ribosomal protein L30 BC095426.1 0.000016 11753694_x_at RPS15A ribosomal protein S15a AB062400.1 0.000017 11755956_x_at POLE3 polymerase (DNA directed), AF070640.1 0.000017 epsilon 3 (p17 subunit) 200063_PM_s_at NPM1 nucleophosmin (nucleolar BC002398.1 0.000017 phosphoprotein B23, numatrin) 11740643_a_at CYP4F8 cytochrome P450, family 4, AF133298.1 0.00002 subfamily F, polypeptide 8 11757305_x_at RPSAP58 ribosomal protein SA BI762726 0.000021 pseudogene 58 11715958_s_at RPL7 ribosomal protein L7 NM_000971.3 0.000023 11719783_at RPS23 ribosomal protein S23 D14530.1 0.000027 11745154_a_at NCL nucleolin BC006516.2 0.000027 11720183_s_at EEF1B2 eukaryotic translation NM_001959.3 0.000029 elongation factor 1 beta 2 11753691_x_at RPL24 ribosomal protein L24 CR456729.1 0.000031 200013_PM_at RPL24 ribosomal protein L24 NM_000986.1 0.000034 11718275_x_at EIF3L eukaryotic translation NM_016091.2 0.000037 initiation factor 3, subunit L 11744326_s_at RPL37 ribosomal protein L37 BC079477.1 0.000037 11757264_s_at RPS3 ribosomal protein S3 BU588459 0.000038 200010_PM_at RPL11 ribosomal protein L11 NM_000975.1 0.000039 11757027_x_at RPL31 ribosomal protein L31 CR600452.1 0.000042 200018_PM_at RPS13 ribosomal protein S13 NM_001017.1 0.000046 11757375_x_at RPS15 ribosomal protein S15 AI625563 0.000047 11754031_s_at CKS1B CDC28 protein kinase BT007196.1 0.000051 regulatory subunit 1B 11715733_a_at NIPSNAP1 nipsnap homolog 1 (C. NM_003634.2 0.000054 elegans) 11740644_x_at CYP4F8 cytochrome P450, family 4, AF133298.1 0.000061 subfamily F, polypeptide 8 11736188_a_at ORMDL3 ORM1-like 3 (S. cerevisiae) NM_139280.1 0.000071 11730527_a_at DAPK2 death-associated protein AF052941.1 0.000088 kinase 2 peroxisomal membrane 11723312_a_at PXMP2 protein 2, 22kDa NM_018663.1 0.000092 11734833_s_at TAF9B TAF9B RNA polymerase II, NM_015975.4 0.000093 TATA box binding protein (TBP)-associated factor, 31kDa 11732205_x_at NAP1L1 nucleosome assembly BX413854 0.000099 protein 1-like 1 11752912_x_at EIF3M eukaryotic translation AK292139.1 0.000099 initiation factor 3, subunit M 11756437_x_at RPS18 ribosomal protein S18 BQ057441 0.0001 11729011_at CDH22 cadherin 22, type 2 NM_021248.1 0.000107 11734329_at TNN tenascin N NM_022093.1 0.000107 11749558_a_at CYP4F8 cytochrome P450, family 4, AK300530.1 0.000119 subfamily F, polypeptide 8 200029_PM_at RPL19 ribosomal protein L19 NM_000981.1 0.000121 11757386_x_at NPM1 nucleophosmin (nucleolar AL563600 0.000122 phosphoprotein B23, numatrin) 11733774_a_at RPL37 ribosomal protein L37 NM_000997.4 0.000123 11729427_a_at GLI1 GLI family zinc finger 1 NM_005269.2 0.000126 11754066_x_at NPM1 nucleophosmin (nucleolar BT007011.1 0.000128 phosphoprotein B23, numatrin) 11715626_a_at RPL11 ribosomal protein L11 NM_000975.2 0.00013 11757489_x_at RPL22 ribosomal protein L22 AW268528 0.00013 11757355_x_at RPL41 ribosomal protein L41 BU958994 0.000138 11752726_x_at GNB2L1 guanine nucleotide binding AY159316.1 0.000144 protein (G protein), beta polypeptide 2-like 1 11728288_a_at KRT15 keratin 15 NM_002275.3 0.000153 11756783_a_at TF transferrin BC045772.1 0.000155 11757331_x_at RPL13A ribosomal protein L13a BF688481 0.000157 11744365_a_at NCRNA00275 non-protein coding RNA 275 AY513722.1 0.000159 11739727_x_at NAP1L1 nucleosome assembly BE965760 0.00016 protein 1-like 1 200074_PM_s_at RPL14 ribosomal protein L14 U16738.1 0.000161 200089_PM_s_at RPL4 ribosomal protein L4 AI953886 0.000162 11734331_a_at TNN tenascin N BC136619.1 0.000163 11757906_x_at RPL10 ribosomal protein L10 AL558950 0.000167 11715645_s_at C22orf28 chromosome 22 open NM_014306.4 0.00018 reading frame 28 11756875_x_at COMMD6 COMM domain containing 6 CR603325.1 0.000184 11722318_a_at EFNB3 ephrin-B3 NM_001406.3 0.000198 11756878_a_at FBL fibrillarin CR593763.1 0.000198 11736721_x_at RPL32 ribosomal protein L32 NM_001007073.1 0.000199 11720184_x_at EEF1B2 eukaryotic translation NM_001959.3 0.0002 elongation factor 1 beta 2 11749786_x_at HNRNPF heterogeneous nuclear AK296696.1 0.000201 ribonucleoprotein F 11717058_x_at RPL5 ribosomal protein L5 NM_000969.3 0.00021 11752911_a_at EIF3M eukaryotic translation AK292139.1 0.000211 initiation factor 3, subunit M 11730790_x_at NPM1 nucleophosmin (nucleolar AK290652.1 0.000213 phosphoprotein B23, numatrin) 11753680_x_at RPL21 ribosomal protein L21 CR457032.1 0.000213 200022_PM_at RPL18 ribosomal protein L18 NM_000979.1 0.000213 200014_PM_s_at HNRNPC heterogeneous nuclear NM_004500.1 0.000215 ribonucleoprotein C (C1/C2) 11742667_x_at RPS28 ribosomal protein S28 NM_001031.4 0.000242 200082_PM_s_at RPS7 ribosomal protein S7 AI805587 0.00025 11728380_x_at NACA2 nascent polypeptide- NM_199290.2 0.000252 associated complex alpha subunit 2 11756140_s_at RPL4 ribosomal protein L4 BX447218 0.000254 11716304_a_at ABHD14B abhydrolase domain NM_032750.2 0.000267 containing 14B 11758357_x_at RPL9 ribosomal protein L9 BF172613 0.000272 11715280_s_at KRT17 keratin 17 g197383031 0.000274 11739813_a_at FZD1 frizzled homolog 1 BF675672 0.000295 (Drosophila) 11721885_s_at CDC42 cell division cycle 42 (GTP NM_001039802.1 0.000307 binding protein, 25kDa) 11720954_s_at RPL30 ribosomal protein L30 NM_000989.2 0.000309 11743688_at GLI2 GLI family zinc finger 2 AB209354.1 0.000312 11720599_s_at SUB1 SUB1 homolog NM_006713.3 0.000315 (S. cerevisiae) 11725875_at WDR66 WD repeat domain 66 NM_144668.4 0.000318 11733496_x_at COMMD6 COMM domain containing 6 AA535445 0.000323 11727795_x_at EIF3E eukaryotic translation NM_001568.2 0.000327 initiation factor 3, subunit E 11756215_x_at UBA52 ubiquitin A-52 residue BU619323 0.000331 ribosomal protein fusion product 1 11757059_x_at RPL36A ribosomal protein L36a CR617894.1 0.000331 200012_PM_x_at RPL21 ribosomal protein L21 NM_000982.1 0.000335 11718344_a_at CNOT7 CCR4-NOT transcription NM_013354.5 0.00035 complex, subunit 7 11717235_s_at RPS7 ribosomal protein S7 NM_001011.3 0.000352 11757113_a_at SNHG1 small nucleolar RNA host 6E836747 0.000365 gene 1 (non-protein coding) 11743679_a_at PTCH1 patched 1 DB299015 0.000374 11744366_a_at NCRNA00275 non-protein coding RNA 275 CR936805.1 0.000378

TABLE 13 Type II Epidermis; Up-regulated GeneTitan_ID Gene Title Public ID p 11729461_a_at CTNS cystinosis, nephropathic NM_001031681.2 0.000105 11737824_a_at STX16 syntaxin 16 NM_001134773.1 0.000753 11731828_at GPC2 glypican 2 NM_152742.1 0.000819 11757259_x_at SCARNA9L small Cajal body-specific NR_023358.1 0.000852 RNA 9-like (retrotransposed) 11728498_a_at SVIL supervillin NM_003174.3 0.001387 11733298_a_at VIPR1 vasoactive intestinal NM_004624.3 0.00166 peptide receptor 1 11754972_s_at BAZ2A bromodomain adjacent to AK127775.1 0.001742 zinc finger domain, 2A 11732899_s_at SULT1AI sulfotransferase family, NM_177528.1 0.001887 cytosolic, 1A, phenol- preferring, member 1 11716708_a_at DDR1 discoidin domain receptor NM_013993.2 0.001925 tyrosine kinase 1 11757623_s_at RNF5 ring finger protein 5 AA923467 0.00209 11715799_s_at BAT2L1 HLA-B associated NM_013318.3 0.002325 transcript 2-like 1 11756190_a_at CLK3 CDC-like kinase 3 CD743118 0.002384 11726634_a_at MYST3 MYST histone NM_001099412.1 0.002824 acetyltransferase (monocytic leukemia) 3 11752331_s_at SULT1A4 sulfotransferase family, BC111011.1 0.002837 cytosolic, 1A, phenol- preferring, member 4 11731093_s_at BRD1 bromodomain containing NM_014577.1 0.002905 1 11744831_a_at RPAIN RPA interacting protein AY775316.1 0.002976 11744173_x_at DNAJC4 DnaJ (Hsp40) homolog, BQ267791 0.00329 subfamily C, member 4 11723546_s_at PLD1 phospholipase D1, BF434088 0.003418 phosphatidylcholine- specific 11732589_a_at ZNF467 zinc finger protein 467 NM_207336.1 0.004153 11721165_a_at KHNYN KH and NYN domain NM_015299.2 0.00435 containing 11730324_s_at SLC38A9 solute carrier family 38, NM_173514.2 0.004534 member 9 11739669_at SS18L1 synovial sarcoma AB014593.1 0.004553 translocation gene on chromosome 18-like 1 11758140_s_at CPSF6 cleavage and BU689332 0.004569 polyadenylation specific factor 6, 68kDa 11721912 at MDM4 Mdm4 p53 binding NM_002393.3 0.004611 protein homolog (mouse) 11744830_x_at NPIPL3 nuclear pore complex AK303166.1 0.004653 interacting protein-like 3 11729100_a_at TTC18 tetratricopeptide repeat NM_145170.3 0.004727 domain 18 11757896_s_at C1orf63 chromosome 1 open R81538 0.004766 reading frame 63 11715976_a_at VGLL4 vestigial like 4 NM_001128219.1 0.004868 (Drosophila) 11729196_a_at STX16 syntaxin 16 6E782754 0.004954 11758055_x_at RGPD8 RANBP2-like and GRIP BQ005433 0.005005 domain containing 8 11721624_s_at WSB1 WD repeat and SOCS box- NM_015626.8 0.005017 containing 1 11720589_s_at PHF21A PHD finger protein 21A BU733437 0.005058 11720895_at SOS1 son of seven less homolog BM970418 0.005891 1 (Drosophila) 11761133_at KDM5C lysine (K)-specific EF613277.1 0.006011 demethylase 5C 11726189_x_at HCFC1R1 host cell factor C1 NM_017885.2 0.006166 regulator 1 (XPO1 dependent) 11721598_a_at EFS embryonal Fyn-associated NM_032459.1 0.006547 substrate 11745431_a_at SVIL supervillin BC092440.1 0.006666 11740447_x_at BTN3A1 butyrophilin, subfamily 3, NM_194441.2 0.006686 member Al 11726515_a_at CLK4 CDC-like kinase 4 AF294429.1 0.006691 11746529_x_at TNFRSF14 tumor necrosis factor BC029848.1 0.006897 receptor superfamily, member 14 (herpesvirus entry mediator) 11759308_s_at MAGI1 membrane associated AL050184.1 0.006929 guanylate kinase, WW and PDZ domain containing 1 11749473_a_at MEF2D myocyte enhancer factor BC040949.1 0.007372 2D 11719128_a_at LMF2 lipase maturation factor 2 NM_033200.1 0.007569 11762365_x_at KIAA0415 KIAA0415 AB007875.1 0.007605 11716129_at IGF2R insulin-like growth factor NM_000876.2 0.007692 2 receptor 11717989_a_at SUN1 Sad1 and UNC84 domain NM_001130965.1 0.007807 containing 1 11755196_a_at CORO6 coronin 6 AK092430.1 0.007848 11755758_s_at NLRC5 NLR family, CARD domain AK090439.1 0.00788 containing 5 11716283_at PAPD7 PAP associated domain NM_006999.3 0.008191 containing 7 11730449_a_at DHRS12 dehydrogenase/reductase NM_024705.1 0.008218 (SDR family) member 12 11718728_a_at ZNF655 zinc finger protein 655 NM_001083956.1 0.008286 11718820_at TSC1 tuberous sclerosis 1 NM_000368.3 0.008558 11729483_a_at KLF8 Kruppel-like factor 8 NM_007250.4 0.008607 11754192_s_at SRSF11 serine/arginine-rich CR614713.1 0.009136 splicing factor 11 11758557_s_at ZFP36L1 zinc finger protein 36, AI758505 0.009209 C3H type-like 1 11723598_x_at MAP2K7 mitogen-activated protein NM_145185.2 0.009355 kinase kinase 7 11754541_a_at CCDC45 coiled-coil domain AW167096 0.00982 containing 45 11757630_s_at HERPUD2 HERPUD family member 2 AA709265 0.010052 11718536_s_at NKTR natural killer-tumor NM_005385.3 0.010291 recognition sequence 11755674_s_at RALGAPAI Ral GTPase activating DQ786317.1 0.010335 protein, alpha subunit 1 (catalytic) 11757591_s_at PAN3 PAN3 poly(A) specific DB314869 0.010593 ribonuclease subunit homolog (S. cerevisiae) 11724312_a_at SH3BP2 SH3-domain binding NM_001145855.1 0.010664 protein 2 11757197_s_at NCRNA00201 non-protein coding RNA NR_026778.1 0.010882 201 11759150_at CNOT4 CCR4-NOT transcription BC035590.1 0.011523 complex, subunit 4 11718939_s_at TNFAIP3 tumor necrosis factor, NM_006290.2 0.011563 alpha-induced protein 3 11738035_s_at RTN4 reticulon 4 AK302741.1 0.011615 11719084_a_at SMARCC2 SWI/SNF related, matrix BF663402 0.011872 associated, actin dependent regulator of chromatin, subfamily c, member 2 11736501_x_at SS18 synovial sarcoma NM_005637.2 0.011911 translocation, chromosome 18 11755811_a_at ZNF266 zinc finger protein 266 AL833503.1 0.012172 11720362_at PHIP pleckstrin homology CR600369.1 0.012352 domain interacting protein 11724758_s_at GPBP1L1 GC-rich promoter binding NM_021639.4 0.012429 protein 1-like 1 solute carrier family 38, 11754821_s_at SLC38AI member 1 AI476037 0.012431 11758907_at ZNF827 zinc finger protein 827 AA031947 0.012473 trinucleotide repeat 11736498_a_at TNRC6B containing 6B NM_015088.2 0.012866 11716010_s_at DYNC1LI2 dynein, cytoplasmic 1, NM_006141.2 0.01317 light intermediate chain 2 11745723_a_at MALAT1 metastasis associated BX538238.1 0.013758 lung adenocarcinoma transcript 1 (non-protein coding) 11758584_s_at STYX serine/threonine/tyrosine N34305 0.013878 interacting protein 11757558_s_at LONRF1 LON peptidase N-terminal BF680438 0.014197 domain and ring finger 1 11750922_x_at AMT aminomethyltransferase AK296177.1 0.0143 11718558_s_at MKRN1 makorin ring finger NM_001145125.1 0.014465 protein 1 11723112_a_at CCDC84 coiled-coil domain containing 84 NM_198489.1 0.014471 11757808_s_at RERE arginine-glutamic acid BM706668 0.014565 dipeptide (RE) repeats 11763191_at PRICKLE3 prickle homolog 3 AK303308.1 0.014697 (Drosophila) 11757821_s_at LDB1 LIM domain binding 1 AW271288 0.014809 11755194_s_at CCN L2 cyclin L2 AK000685.1 0.014941 11720122_at GIGYF1 GRB10 interacting GYF NM_022574.4 0.014956 protein 1 hypothetical protein 11763351_at LOC286052 LOC286052 CK819455 0.014973 11722752_a_at C14orf43 chromosome 14 open NM_194278.3 0.015053 reading frame 43 11757958_s_at POGZ pogo transposable AI374931 0.015063 element with ZNF domain 11734056_at PTGR2 prostaglandin reductase 2 NM_152444.2 0.015377 11722715_at STK35 serine/threonine kinase NM_080836.2 0.015673 35 11722134_a_at TNFRSF25 tumor necrosis factor NM_148965.1 0.015688 receptor superfamily, member 25 11715192_s_at C7orf46 chromosome 7 open g188219621 0.015694 reading frame 46 11720795_s_at NUPL1 nucleoporin like 1 NM_014089.3 0.016154 11729510_a_at WDR33 WD repeat domain 33 NM_001006623.1 0.016469 11724066_s_at HCFC1R1 host cell factor C1 NM_001002018.1 0.016491 regulator 1 (XPO1 dependent) 11718534_at NKTR natural killer-tumor AI361805 0.016611 recognition sequence 11741625_a_at SLC22A23 solute carrier family 22, NM_021945.5 0.016631 member 23 11722305_at ARHGAP23 Rho GTPase activating NM_020876.1 0.01665 protein 23 11764248_s_at LDLRAD3 low density lipoprotein AW043782 0.016656 receptor class A domain containing 3

TABLE 14 Type II Epidermis; Down-regulated GeneTitan_ID Gene Title Public ID p homeodomain interacting 11719408_a_at HIPK2 protein kinase 2 BM679184 0.000155 11739028_s_at CLTC clathrin, heavy chain (Hc) BX395378 0.00019 11747337_x_at EIF3I eukaryotic translation U36764.1 0.000378 initiation factor 3, subunit I TBC1 domain family, 11749845_a_at TBC1D22A member 22A AK301445.1 0.000577 sec1 family domain 11723960_at SCFD2 containing 2 BC032453.1 0.000699 proteasome (prosome, macropain) subunit, beta 11717105_a_at PSMB5 type, 5 NM_001144932.1 0.00071 proteasome (prosome, macropain) 26S subunit, 11716545_x_at PSMC1 ATPase, 1 NM_002802.2 0.00077 1-acylglycerol-3-phosphate O-acyltransferase 5 (lysophosphatidic acid 11730754_s_at AGPAT5 acyltransferase, epsilon) CB306609 0.00077 11750994_a_at SYAP1 synapse associated protein 1 AK295322.1 0.000788 11754977_x_at CTSB cathepsin B CR614817.1 0.000808 ATPase, H+ transporting, lysosomal 9kDa, V0 subunit 200096_PM_s_at ATP6V0E1 e1 AI862255 0.000889 serpin peptidase inhibitor, clade B (ovalbumin), 11738899_a_at SERPINB12 member 12 NM_080474.1 0.000913 G-rich RNA sequence 11747533_a_at GRSF1 binding factor 1 AK298883.1 0.000973 proteasome (prosome, macropain) 26S subunit, 11733918_a_at PSMD14 non-ATPase, 14 BC066336.1 0.001091 11753572_a_at TMEM85 transmembrane protein 85 AY336092.1 0.001121 glucosidase, alpha; neutral 11738988_a_at GANAB AB AK302752.1 0.001195 golgi reassembly stacking 11751303_s_at GORASP2 protein 2, 55kDa AK293640.1 0.001298 mitochondrial ribosomal 11719482_a_at MRPL21 protein L21 NM_181515.1 0.001366 proteasome (prosome, macropain) subunit, beta 11715943_x_at PSMB1 type, 1 NM_002793.3 0.001475 11749935_s_at TPM3 tropomyosin 3 AK298678.1 0.001523 succinate dehydrogenase complex, subunit C, integral 11736759_s_at SDHC membrane protein, 15kDa AB211234.1 0.001635 tyrosine 3- monooxygenase/tryptophan 5-monooxygenase activation 11731397_a_at YWHAB protein, beta polypeptide AI866370 0.001646 mediator complex subunit 11751557_s_at MED27 27 AK298436.1 0.001692 transmembrane BAX 11747146_s_at TMBIM6 inhibitor motif containing 6 AK304577.1 0.001727 N-ethylmaleimide-sensitive 11716972_s_at NSF factor NM_006178.2 0.001747 proteasome (prosome, macropain) subunit, beta 11754009_a_at PSMB5 type, 5 BT006777.1 0.001747 mitochondrial ribosomal 11717459_a_at MRPL39 protein L39 NM_017446.3 0.001824 mitochondrial ribosomal 11742273_a_at MRPL33 protein L33 AF420602.1 0.001825 11739599_a_at ZNF398 zinc finger protein 398 BU736496 0.001891 11747534_a_at RSU1 Ras suppressor protein 1 BC008691.1 0.002033 nucleosome assembly 11747507_x_at NAP1L4 protein 1-like 4 AK316548.1 0.002069 nucleosome assembly 11747506_a_at NAP1L4 protein 1-like 4 AK316548.1 0.002092 phosphatidylinositol binding 11748665_a_at PICALM clathrin assembly protein AK300275.1 0.002111 peptidylprolyl isomerase 11718035_at PPIL1 (cyclophilin)-like 1 AF151882.1 0.002124 11751336_x_at MKRN1 makorin ring finger protein 1 AK297361.1 0.002138 activating signal cointegrator 1 complex 11720264_at ASCC3 subunit 3 NM_006828.2 0.002301 11725037_a_at 5EC23IP 5EC23 interacting protein AK000698.1 0.002534 mitochondrial ribosomal 11715772_x_at MRPL13 protein L13 NM_014078.4 0.002536 prolyl 4-hydroxylase, beta 11716173_a_at P4HB polypeptide AK296206.1 0.002544 proteasome (prosome, macropain) subunit, beta 11717106_x_at PSMB5 type, 5 NM_001144932.1 0.002579 ras homolog gene family, 200059_PM_s_at RHOA member A BC001360.1 0.002604 11746655_a_at ACAAI acetyl-CoA acyltransferase 1 AK303251.1 0.002689 ATP synthase, H+ transporting, mitochondrial Fo complex, subunit C3 11739201_a_at ATP5G3 (subunit 9) 6E736890 0.002714 ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, 11728637_a_at ATP5AI cardiac muscle NM_001001937.1 0.002718 BCL2-like 10 (apoptosis 11756013_a_at BCL2L10 facilitator) BC143227.1 0.002726 proteasome (prosome, macropain) subunit, beta 11754271_a_at PSMB4 type, 4 BM849884 0.002734 proteasome (prosome, macropain) 26S subunit, 11731415_a_at PSMD6 non-ATPase, 6 NM_014814.1 0.002807 succinate dehydrogenase complex, subunit C, integral 11715840_s_at SDHC membrane protein, 15kDa BC020808.1 0.002866 succinate-CoA ligase, ADP- 11755266_x_at SUCLA2 forming, beta subunit AK001458.1 0.003146 sec1 family domain 11750876_a_at SCFD1 containing 1 AK301406.1 0.003266 SWI/SNF related, matrix associated, actin dependent regulator of chromatin, 11752770_a_at SMARCE1 subfamily e, member 1 AK294666.1 0.003311 SERPINE1 mRNA binding 11758800_x_at SERBP1 protein 1 AF151813.1 0.003464 11751835_a_at LTV1 LTV1 homolog (S. cerevisiae) AY326463.1 0.003472 11758319_x_at UBC ubiquitin C BF672950 0.003583 11729168_x_at DCTD dCMP deaminase BC001286.1 0.003633 NADH dehydrogenase (ubiquinone) 1 beta 11717159_a_at NDUFB3 subcomplex, 3, 12kDa NM_002491.2 0.003697 phenylalanyl-tRNA 11744181_a_at FARS2 synthetase 2, mitochondrial BG192794 0.003735 ADP-ribosylation factor-like 11751412_x_at ARL1 1 AK301701.1 0.003805 proteasome (prosome, macropain) subunit, alpha 11734682_a_at PSMA7 type, 7 NM_002792.2 0.003878 11750059_a_at MLX MAX-like protein X AK296114.1 0.003884 zinc finger, HIT-type 11715718_a_at ZNHIT1 containing 1 NM_006349.2 0.003894 REX2, RNA exonuclease 2 11751360_x_at REX02 homolog (S. cerevisiae) BC107887.1 0.003907 phosphoserine 11747349_s_at PSAT1 aminotransferase 1 BT006840.1 0.003982 mitochondrial ribosomal 11763975_a_at MRPS11 protein S11 DB346141 0.004073 11716381_x_at BRP44 brain protein 44 NM_015415.2 0.00408 transmembrane emp24 protein transport domain 11751523_a_at TMED5 containing 5 AK293308.1 0.004086 small nuclear ribonucleoprotein 11753974_s_at SNRPG polypeptide G CR456918.1 0.004159 penta-EF-hand domain 11732216_s_at PEF1 containing 1 CR542139.1 0.004221 calpain 2, (m/II) large 11718978_x_at CAPN2 subunit BC021303.2 0.004238 ATPase, H+ transporting, lysosomal 38kDa, V0 subunit 11763422_a_at ATP6V0D1 dl BX397389 0.004241 11754060_a_at DAD1 defender against cell death CR542204.1 0.004283 signal peptidase complex subunit 1 homolog (S. 11743905_a_at SPCS1 cerevisiae) 6E782150 0.004361 proteasome (prosome, macropain) subunit, beta 11715732_at PSMB3 type, 3 NM_002795.2 0.004369 Yip1 domain family, member 11751505_a_at YIPF1 1 AK300240.1 0.004409 chromosome 11 open 11742925_a_at C11orf59 reading frame 59 CR457247.1 0.004503 S-phase kinase-associated 11715417_s_at SKP1 protein 1 BC020798.1 0.004548 phosphoglycerate mutase 1 11750438_x_at PGAM1 (brain) AK296619.1 0.004563 11752939_x_at PGK1 phosphoglycerate kinase 1 AK298855.1 0.004575 cell division cycle 123 11723478_s_at CDC123 homolog (S. cerevisiae) NM_006023.2 0.004585 retinol dehydrogenase 11 11743034_x_at RDH11 (all-trans/9-cis/11-cis) AK289427.1 0.004593 VAMP (vesicle-associated membrane protein)- 11754086_x_at VAPA associated protein A, 33kDa BT019618.1 0.004657 heterogeneous nuclear ribonucleoprotein D (AU- rich element RNA binding 11749303_s_at HNRNPD protein 1, 37kDa) AK300149.1 0.004671 proteasome (prosome, macropain) 26S subunit, 11753142_a_at PSMD11 non-ATPase, 11 AK300342.1 0.004673 11747719_a_at KIAA0391 KIAA0391 AK304066.1 0.004719 CWF19-like 1, cell cycle 11748974_s_at CWF19L1 control (S. pombe) AK295303.1 0.004746 ATPase, H+ transporting, lysosomal 56/58kDa, V1 11751133_a_at ATP6V1B2 subunit B2 AK298819.1 0.004759 inner membrane protein, 11715552_a_at IMMT mitochondrial NM_006839.2 0.004766 11747365_a_at QARS glutaminyl-tRNA synthetase AK302867.1 0.004855 succinate dehydrogenase complex, subunit D, integral 11758248_s_at SDHD membrane protein BF696015 0.004909 eukaryotic translation 11753592_x_at EEF1G elongation factor 1 gamma AK299876.1 0.004961 NADH dehydrogenase (ubiquinone) 1 alpha 11715369_s_at NDUFA4 subcomplex, 4, 9kDa BC105295.1 0.005125 11715499_x_at CBX3 chromobox homolog 3 U26312.1 0.005141 ATP synthase, H+ 11715874_s_at ATP5H transporting, mitochondrial NM_006356.2 0.005143 Fo complex, subunit d succinate dehydrogenase complex, subunit D, integral 11758311_s_at SDHD membrane protein BF697775 0.005199 11715883_x_at DAP3 death associated protein 3 NM_004632.2 0.005251 poly(A) binding protein, cytoplasmic 4 (inducible 11754030_a_at PABPC4 form) BC118568.1 0.00537 exocyst complex component 11749682_s_at EXOC5 5 AK303531.1 0.005371 tetratricopeptide repeat 11718142_a_at TTC27 domain 27 NM_017735.3 0.005424 thioredoxin domain 11754067_a_at TXNDC9 containing 9 CR456935.1 0.005482 aldo-keto reductase family 1, member A1 (aldehyde 11716509_a_at AKR1A1 reductase) NM_006066.2 0.005483

TABLE 15 Type II Dermis; Up-regulated GeneTitan_ID Gene Title Public ID p 11715351_at COL1AI collagen, type I, alpha 1 NM_000088.3 0.000139 11715352_x_at COL1AI collagen, type I, alpha 1 NM_000088.3 0.000375 11734105 a_at PNMAL2 PNMA-like 2 AB033009.1 0.001583 11763844_s_at UBXN6 UBX domain protein 6 CR590857.1 0.001819 11756896_a_at COL6A6 collagen, type VI, alpha 6 AL713792.1 0.002188 chromosome 15 open reading 11715284_x_at C15orf40 frame 40 g237858663 0.003648 platelet-derived growth factor receptor, beta 11715852_at PDGFRB polypeptide NM_002609.3 0.005919 phosphatidylinositol-5- phosphate 4-kinase, type II, 11715888_s_at PIP4K2B beta NM_003559.4 0.006199 chromosome 5 open reading 11724481_a_at C5orf13 frame 13 NM_004772.2 0.006404 11758388_s_at LHX8 LIM homeobox 8 DB302169 0.006562 G protein-coupled receptor 11727836_a_at GPR78 78 NM_001014447.1 0.007388 family with sequence 11729827_at FAM110B similarity 110, member B BCO24294.1 0.009523 family with sequence 11744562_x_at FAM176B similarity 176, member B BC071697.1 0.009813 11725867_s_at EBF3 early B-cell factor 3 NM_001005463.1 0.009883 progestin and adipoQ 11749069_a_at PAQR4 receptor family member IV AK295348.1 0.010054 corticotropin releasing 11723068_at CRHBP hormone binding protein NM_001882.3 0.011101 fibronectin type III domain 11723174_a_at FNDC1 containing 1 NM_032532.2 0.011173 11717274_s_at COL5AI collagen, type V, alpha 1 BQ007762 0.012177 calcium/calmodulin- dependent protein kinase 11729541_a_at CAMKK2 kinase 2, beta AB081337.1 0.012395 11724848_a_at DIXDC1 DIX domain containing 1 DB358954 0.013451 11726830_at ANTXR1 anthrax toxin receptor 1 NM_018153.3 0.013945 transforming growth factor, 11727296_s_at TGFB3 beta 3 NM_003239.2 0.014199 exonuclease 3′-5′ domain 11759905_a_at EXD3 containing 3 BC110879.1 0.01495 transcription factor 7-like 1 11721372_at TCF7L1 (T-cell specific, HMG-box) NM_031283.1 0.01515 fibroblast activation protein, 11755955_a_at alpha AL832166.1 FAP 0.015704 matrix metallopeptidase 14 11725989_x_at MMP14 (membrane-inserted) NM_004995.2 0.019095 11717272_at COL5AI collagen, type V, alpha 1 AB371583.1 0.020092 chromosome 19 open reading 11739544_a_at C19orf12 frame 12 BX328123 0.021 C-type lectin domain family 3, 11727867_a_at CLEC3B member B NM_003278.2 0.024069 calcium/calmodulin- dependent protein kinase 11731645_a_at CAMKK2 kinase 2, beta BCO26060.2 0.024228 aldo-keto reductase family 1, member C2 (dihydrodiol dehydrogenase 2; bile acid binding protein; 3-alpha hydroxysteroid 11729101_a_at AKR1C2 dehydrogenase, type III) NM_205845.1 0.024814 hairy and enhancer of split 4 11726474_a_at HES4 (Drosophila) NM_021170.3 0.025047 11725224_a_at ZNF193 zinc finger protein 193 NM_006299.3 0.026826 11715350_a_at COL1AI collagen, type I, alpha 1 BC036531.2 0.026857 matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV 11741377_a_at MMP2 collagenase) NM_001127891.1 0.02687 NYN domain and retroviral 11722292_a_at NYNRIN integrase containing NM_025081.2 0.027121 11730404_at MEX3B mex-3 homolog B (C. elegans) NM_032246.3 0.027161 thyroid hormone receptor, alpha (erythroblastic leukemia viral (v-erb-a) 11759126_a_at THRA oncogene homolog, avian) CB054873 0.02738 11744348_x_at COL6A2 collagen, type VI, alpha 2 BC002484.2 0.027551 11720845_a_at CD248 CD248 molecule, endosialin NM_020404.2 0.02836 11720372_at TESC tescalcin NM_017899.2 0.028677 syntrophin, alpha 1 (dystroph in-associated protein Al, 59kDa, acidic 11752890_a_at SNTAI component) AK301800.1 0.029777 11717273_at COL5AI collagen, type V, alpha 1 BQ007762 0.030174 aldehyde dehydrogenase 1 11754184_a_at ALDH1A3 family, member A3 BX538027.1 0.030605 TRIO and F-actin binding 11727155_a_at TRIOBP protein NM_007032.5 0.030721 11727031_a_at SQSTM1 sequestosome 1 NM_003900.4 0.031363 11720846_at CD248 CD248 molecule, endosialin NM_020404.2 0.031411 triple functional domain 11761938_a_at TRIO (PTPRF interacting) AB115332.1 0.032433 11718096_a_at MEF2A myocyte enhancer factor 2A BC013437.2 0.03254 neuro-oncological ventral 11734906_a_at NOVAI antigen 1 NM_002515.2 0.032886 R-spondin 3 homolog 11724619_at RSPO3 (Xenopus laevis) NM_032784.3 0.033442 shisa homolog 3 (Xenopus 11726188_at SHISA3 laevis) NM_001080505.1 0.033466 glutathione peroxidase 8 11729644_a_at GPX8 (putative) AK074216.1 0.033979 adrenergic, alpha-2C-, 11756359_s_at ADRA2C receptor CR590957.1 0.034296 11747064_x_at ANXAll annexin All AK301047.1 0.034459 chromosome 16 open reading 11732785_a_at Cl6orf45 frame 45 NM_001142469.1 0.035589 microtubule-associated 11727030_s_at MAP1A protein 1A NM_002373.5 0.036125 sema domain, immunoglobulin domain (Ig), short basic domain, secreted, 11748738_a_at SEMA3E (semaphorin) 3E AK303925.1 0.036605 leucine rich repeat containing 11721995_a_at LRRC32 32 NM_001128922.1 0.038953 host cell factor Cl regulator 1 11726189_x_at HCFC1R1 (XPO1 dependent) NM_017885.2 0.039807 RGM domain family, member 11754792_a_at RGMA A AK125204.1 0.040435 protein phosphatase 1, regulatory (inhibitor) subunit 11717123_a_at PPP1R12B 12B NM_032105.1 0.043242 triple functional domain 11724260_a_at TRIO (PTPRF interacting) AF091395.1 0.044219 11759962_at TPRKB TP53RK binding protein AY643713.1 0.045288 11733167_at LRRN4CL LRRN4 C-terminal like BC053902.1 0.045766 trinucleotide repeat 11721703_s_at TN RC18 containing 18 NM_001080495.2 0.045935 ATPase, aminophospholipid transporter (APLT), class I, 11725568_a_at ATP8AI type 8A, member 1 NM_001105529.1 0.046173 membrane metallo- 11741562_a_at MME endopeptidase NM_007287.2 0.046616 pleiomorphic adenoma gene- 11745820_s_at PLAGL1 like 1 BQ026948 0.046856 C-type lectin domain family 11743696_at CLEC14A 14, member A CA412481 0.047788 11720277_a_at OLFML2A olfactomedin-like 2A NM_182487.2 0.049248

TABLE 16 Type II Dermis; Down-regulated GeneTitan_ID Gene Title Public ID p microtubule-associated 11749128_x_at MAP7 protein 7 AK299355.1 0.000033 eukaryotic translation 11720184_x_at EEF162 elongation factor 1 beta 2 NM_001959.3 0.000101 sulfotransferase family 1E, estrogen-preferring, member 11724155_at SULT1E1 1 U08098.1 0.000109 200062_PM_s_at RPL30 ribosomal protein L30 L05095.1 0.000123 11742734_s_at WDR3 WD repeat domain 3 AK292438.1 0.000139 nucleosome assembly protein 11732205_x_at NAP1L1 1-like 1 BX413854 0.000141 11743604_s_at RRM1 ribonucleotide reductase M1 6E618815 0.000164 heterogeneous nuclear 11723197_at HNRNPA3 ribonucleoprotein A3 BX434302 0.000178 ATP-binding cassette, sub- 11732684_a_at ABCAI2 family A (ABC1), member 12 AF418105.1 0.000237 discs, large homolog 1 11739308_s_at DLG1 (Drosophila) BM681931 0.000254 11725875_at WDR66 WD repeat domain 66 NM_144668.4 0.000271 200082_PM_s_at RPS7 ribosomal protein S7 AI805587 0.00036 200081_PM_s_at RPS6 ribosomal protein S6 6E741754 0.000371 nucleophosmin (nucleolar phosphoprotein 623, 200063_PM_s_at NPM1 numatrin) BC002398.1 0.000404 11757356_x_at RPL30 ribosomal protein L30 BM855760 0.000405 11719783_at RPS23 ribosomal protein S23 D14530.1 0.000416 sulfotransferase family 1E, estrogen-preferring, member 11724156_at SULT1E1 1 NM_005420.2 0.000461 eukaryotic translation 11720183_s_at EEF162 elongation factor 1 beta 2 NM_001959.3 0.000477 11745154_a_at NCL nucleolin BC006516.2 0.000485 11715958_s_at RPL7 ribosomal protein L7 NM_000971.3 0.000491 11749558_a_at CYP4F8 cytochrome P450, family 4, AK300530.1 0.000497 subfamily F, polypeptide 8 11755203_x_at RPL21 ribosomal protein L21 BX647669.1 0.000511 11728022_a_at TMEM45A transmembrane protein 45A NM_018004.1 0.000533 eukaryotic translation 11718273_a_at EIF3L initiation factor 3, subunit L NM_016091.2 0.000658 protein kinase, DNA-activated, 11757399_s_at PRKDC catalytic polypeptide AV760328 0.000698 chromosome 10 open reading 11736055_at C10orf32 frame 32 BG696280 0.000732 steroid-5-alpha-reductase, alpha polypeptide 1 (3-oxo-5 alpha-steroid delta 4- 11749267_a_at SRD5A1 dehydrogenase alpha 1) AK315996.1 0.000744 transmembrane 9 superfamily 11716946_s_at TM9SF3 member 3 AF269150.1 0.000758 11726461_a_at PDCD2 programmed cell death 2 NM_144781.1 0.000759 11763318_s_at CSNK1A1 casein kinase 1, alpha 1 BC040473.1 0.000792 eukaryotic translation 11727794_s_at EIF3E initiation factor 3, subunit E NM_001568.2 0.000802 phosphoinositide-3-kinase, 11731690_a_at PIK3C2G class 2, gamma polypeptide NM 004570.4 0.000846 11748052_x_at E124 etoposide induced 2.4 mRNA AK316539.1 0.000851 11743094_at SPRR4 small proline-rich protein 4 BC069445.1 0.000998 11717236_x_at RPS7 ribosomal protein S7 NM_001011.3 0.001062 11718719_at K1AAI797 K1AAI797 NM_017794.3 0.001102 transcription elongation factor 11752908_a_at TCEA1 A (SII), 1 AK297729.1 0.001141 11753659_x_at RPL30 ribosomal protein L30 BC095426.1 0.001161 11717058_x_at RPL5 ribosomal protein L5 NM_000969.3 0.001214 chromosome 10 open reading 11736057_s_at C10orf32 frame 32 BU685637 0.00123 phosphoserine 11742991_a_at PSAT1 aminotransferase 1 AK295222.1 0.001244 11758357_x_at RPL9 ribosomal protein L9 BF172613 0.00126 200010_PM_at RPL11 ribosomal protein L11 NM_000975.1 0.001282 transcription factor AP-2 gamma (activating enhancer 11749776_a_at TFAP2C binding protein 2 gamma) AK301572.1 0.001283 serine peptidase inhibitor, 11727995_a_at SPINK5 Kazal type 5 DQ149928.1 0.001284 chromosome 14 open reading 11722185_at C14orf147 frame 147 NM_138288.3 0.001309 eukaryotic translation 11750883_a_at EIF2A initiation factor 2A, 65kDa AF109358.1 0.001327 200017_PM_at RPS27A ribosomal protein S27a NM_002954.1 0.001414 cytochrome P450, family 4, 11749559_x_at CYP4F8 subfamily F, polypeptide 8 AK300530.1 0.001416 11722308_a_at TP63 tumor protein p63 NM_003722.4 0.001433 nucleophosmin (nucleolar phosphoprotein 623, 11754066_x_at NPM1 numatrin) BT007011.1 0.001448 11743603_a_at RRM1 ribonucleotide reductase M1 6E618815 0.001462 serine peptidase inhibitor, 11754963_a_at SPINK5 Kazal type 5 AK301660.1 0.001558 hydroxysteroid (17-beta) 11747333_a_at HSD1764 dehydrogenase 4 AK295440.1 0.001571 nucleophosmin (nucleolar phosphoprotein 623, 11757386_x_at NPM1 numatrin) AL563600 0.00169 cytochrome P450, family 4, 11751437_a_at CYP4F8 subfamily F, polypeptide 8 AK300539.1 0.001708 11726258_at RNF141 ring finger protein 141 BX503543 0.001716 11720766_a_at METTL9 methyltransferase like 9 AK074529.1 0.001743 coiled-coil domain containing 11743729_at CCDC47 47 AL575693 0.001771 heterogeneous nuclear 11749786_x_at HNRNPF ribonucleoprotein F AK296696.1 0.001879 eukaryotic translation 11729152_a_at EIF3M initiation factor 3, subunit M NM_006360.3 0.001895 UDP glucuronosyltransferase 2 11737634_a_at UGT2A1 family, polypeptide Al NM_006798.2 0.001986 11748044_a_at SCEL sciellin BCO20726.1 0.002038 eukaryotic translation 11752912_x_at EIF3M initiation factor 3, subunit M AK292139.1 0.002066 11736309_a_at CSNK1A1 casein kinase 1, alpha 1 L37042.1 0.002093 11727421_a_at CANX calnexin C6243867 0.002106 11727425_s_at CANX cal nexin M94859.1 0.002182 ribosomal protein SA 11757305_x_at RPSAP58 pseudogene 58 61762726 0.002211 transmembrane emp24 200087_PM_s_at TMED2 domain trafficking protein 2 AK024976.1 0.002219 RAB11A, member RAS 11718030_at RAB11A oncogene family NM_004663.3 0.002258 11742887_a_at BAGS BCL2-associated athanogene 5 6Q008934 0.00227 11746199_a_at METTL9 methyltransferase like 9 AA524199 0.002298 phosphoserine 11756182_s_at PSAT1 aminotransferase 1 AAI73918 0.002312 11727658_s_at KLK10 kallikrein-related peptidase 10 AF024605.1 0.002335 nucleosome assembly protein 11732204_a_at NAP1L1 1-like 1 BX413854 0.002341 11744334_x_at RPS17 ribosomal protein S17 BC071928.1 0.002403 DIM1 dimethyladenosine transferase 1-like (S. 11748536_a_at DIMT1L cerevisiae) BC002841.2 0.002406 11745155_s_at NCL nucleolin BC006516.2 0.002458 nucleophosmin (nucleolar phosphoprotein B23, 11730790_x_at NPM1 numatrin) AK290652.1 0.002509 cytochrome P450, family 4, 11740643_a_at CYP4F8 subfamily F, polypeptide 8 AF133298.1 0.002616 ATPase, Ca++ transporting, 11755057_s_at ATP2C1 type 2C, member 1 AB037768.1 0.002687 phosphoserine 11742992_s_at PSAT1 aminotransferase 1 AK295222.1 0.002716 11757424_x_at RPL37 ribosomal protein L37 F34903 0.002789 peroxisomal membrane 11723312_a_at PXMP2 protein 2, 22kDa NM_018663.1 0.002819 11756137_x_at BTF3 basic transcription factor 3 CA772090 0.002824 11754054_x_at RPL3 ribosomal protein L3 L22453.1 0.002905 11750667_a_at RRM1 ribonucleotide reductase M1 AK297988.1 0.00292 RAB11A, member RAS 11751326_a_at RAB11A oncogene family AK300008.1 0.002998 11749040_a_at PGM2 phosphoglucomutase 2 AK297752.1 0.003008 11715376_a_at RPS11 ribosomal protein S11 NM_001015.3 0.003035 DEAD (Asp-Glu-Ala-Asp) box 11715849_a_at DDX47 polypeptide 47 NM_016355.3 0.003047 tRNA-yW synthesizing protein 11726829_at TYW1B 1 homolog B (S. cerevisiae) NM_001145440.1 0.003195 chromosome 14 open reading 11756267_x_at C14orf166 frame 166 BX349547 0.003217 11756437_x_at RPS18 ribosomal protein S18 BQ057441 0.003368 11715648_x_at ADIPOR1 adiponectin receptor 1 AY424279.1 0.003389 prostaglandin E synthase 3 11747662_x_at PTGES3 (cytosolic) AK298147.1 0.003391 solute carrier family 39 (zinc 11749546_a_at 5LC39A6 transporter), member 6 AK301539.1 0.003463 protein disulfide isomerase 11745720_s_at PDIA6 family A, member 6 D49489.1 0.00351 200074_PM_s_at RPL14 ribosomal protein L14 U16738.1 0.003587 phosphogluconate 11746023_a_at PGD dehydrogenase AK304423.1 0.003624

TABLE 17 Type III Epidermis; Up-regulated GeneTitan_ID Gene Title Public ID p 11725675_a_at RORA RAR-related orphan receptor A AA034012 0.00064 11732366_a_at SCAPER S-phase cyclin A-associated BC015212.1 0.001096 protein in the ER 11739639_at CDK12 cyclin-dependent kinase 12 AW968504 0.001477 11745215_a_at KBTBD4 kelch repeat and BTB (POZ) CR457270.1 0.001496 domain containing 4 11730873_a_at RASSF5 Ras association (RalGDS/AF-6) NM_182665.2 0.001694 domain family member 5 11718966_at NUFIP2 nuclear fragile X mental BU533767 0.001959 retardation protein interacting protein 2 11718513_x_at TSPAN14 tetraspanin 14 NM_030927.2 0.002425 11744585_a_at ATRN attractin AK302730.1 0.003547 11755420_a_at KDM4B lysine (K)-specific demethylase 4B AK126854.1 0.003701 11720895_at SOS1 son of sevenless homolog 1 BM970418 0.003776 (Drosophila) 11734873_a_at SCAPER S-phase cyclin A-associated NM_020843.2 0.00429 protein in the ER 11759897_x_at OFD1 oral-facial-digital syndrome 1 BC042830.1 0.004666 11754251_a_at USP36 ubiquitin specific peptidase 36 AK022913.1 0.005023 11736059_a_at KIF5B kinesin family member 5B BC065267.1 0.005318 11754824_a_at HSPC159 galectin-related protein DB323149 0.005358 11757991_s_at ANKRD12 ankyrin repeat domain 12 AA399583 0.006238 11744468_at SYNCRIP synaptotagmin binding, AK056188.1 0.007087 cytoplasmic RNA interacting protein 11754992_a_at CHD1 chromodomain helicase DNA BE535223 0.007613 binding protein 1 11716283_at PAPD7 PAP associated domain NM_006999.3 0.00802 containing 7 11722973_s_at FOXK1 forkhead box K1 NM_001037165.1 0.008695 11748149_a_at FNBP1 formin binding protein 1 AK293743.1 0.009063 11722125_a_at C3orf19 chromosome 3 open reading AL526467 0.009976 frame 19 11720007_a_at STEAP4 STEAP family member 4 NM_024636.2 0.011197 11731506_a_at RAD23B RAD23 homolog B (S. cerevisiae) NM_002874.3 0.011526 11724759_s_at CALM1 calmodulin 1 (phosphorylase NM_006888.3 0.011622 kinase, delta) 11718161_at KLF13 Kruppel-like factor 13 AF132599.1 0.012064 11726305_at C10orf84 chromosome 10 open reading BC023577.2 0.012188 frame 84 11723962_at KIAAI143 KIAAI143 BC008468.1 0.012597 11740956_x_at PLEKHN1 pleckstrin homology domain NM_032129.1 0.013256 containing, family N member 1 11722305_at ARHGAP23 Rho GTPase activating protein 23 NM_020876.1 0.01353 family with sequence similarity 11726022_a_at FAM177AI 177, member AI BC029559.1 0.01364 11754010_x_at GOLGA2 golgin A2 BT007248.1 0.013918 11729523_a_at NLRC5 NLR family, CARD domain NM_032206.3 0.014235 containing 5 11723113_a_at CENPC1 centromere protein C 1 BC041117.1 0.0145 UPF1 regulator of nonsense 11754616_a_at UPF1 transcripts homolog (yeast) AI690963 0.014647 11722291_s_at ZBTB43 zinc finger and BTB domain AI745225 0.014686 containing 43 11739805_a_at RASAL2 RAS protein activator like 2 AK075169.1 0.014761 11723502_at PRLR prolactin receptor AI435838 0.014945 11726244_a_at RORA RAR-related orphan receptor A U04898.1 0.01551 11736104_a_at ZNF750 zinc finger protein 750 BC109037.1 0.015749 11723184_x_at CNOT6L CCR4-NOTt ranscription complex, BQ025327 0.015988 subunit 6-like 11755058_a_at BAZ1A bromodomain adjacent to zinc BC020636.1 0.016704 finger domain, 1A 11759600_at SFRS18 Splicing factor, arginine/serine- AK027751.1 0.016758 rich 18 11744829_s_at HLA-E major histocompatibility AK296822.1 0.016828 complex, class I, E guanylate binding protein 2, 11719447_s_at GBP2 interferon-inducible BC073163.1 0.016832 11720541_at HSPC159 galectin-related protein NM_014181.2 0.017088 pleckstrin and Sec7 domain 11719028_a_at PSD3 containing 3 DB314358 0.017377 11754462_a_at RSPRY1 ring finger and SPRY domain AU253443 0.017904 containing 1 11725676_a_at RORA RAR-related orphan receptor A NM_002943.3 0.018226 11722290_a_at ZBTB43 zinc finger and BTB domain AI745225 0.018329 containing 43 11723821_a_at SMURF2 SMAD specific E3 ubiquitin AY014180.1 0.018461 protein ligase 2 11720111_at SNTB2 syntrophin, beta 2 (dystrophin- BC036429.1 0.018669 associated protein AI, 59kDa, basic component 2) 11736432_x_at PPP4R2 protein phosphatase 4, BC128136.1 0.01869 regulatory subunit 2 11744000_a_at NFKBIA nuclear factor of kappa light BX367826 0.018695 polypeptide gene enhancer in B- cells inhibitor, alpha 11759512_x_at CWC25 CWC25 spliceosome-associated CR748127 0.018768 protein homolog (S. cerevisiae) 11716095_s_at KLF6 Kruppel-like factor 6 CD366698 0.019227 11754447_a_at RPS6KA5 ribosomal protein S6 kinase, BM968829 0.019584 90kDa, polypeptide 5 11719085_a_at SMARCC2 SWI/SNF related, matrix AL544435 0.01997 associated, actin dependent regulator of chromatin, subfamily c, member 2 11727506_x_at RAB21 RAB21, member RAS oncogene BC021901.1 0.020551 family 11720276_s_at TREX1 three prime repair exonuclease 1 NM_016381.3 0.020671 11724549_a_at RSBN1 round spermatid basic protein 1 AK292552.1 0.020675 11731645_a_at CAMKK2 calcium/calmodulin-dependent BC026060.2 0.020762 protein kinase kinase 2, beta 11737413_at MICALCL MICAL C-terminal like NM_032867.2_ 0.020903 11715938_x_at KHDRBS1 KH domain containing, RNA BC000717.1 0.021833 binding, signal transduction associated 1 11747192_x_at NFIC nuclear factor I/C (CCAAT-binding AK289885.1 0.022039 transcription factor) 11729396_a_at NEK1 NIMA (never in mitosis gene a)- Z25431.1 0.022138 related kinase 1 11727064_a_at ANKRD11 ankyrin repeat domain 11 BU674634 0.022386 11752626_a_at PBX1 pre-B-cell leukemia homeobox 1 AK299673.1 0.022397 11721119_a_at ANKHD1- ANKHD1-EIF4EBP3 readthrough AF217646.1 0.022562 EIF4EBP3 11743648_a_at DCAF6 DDB1 and CUL4 associated factor BF672818 0.022893 6 11740362_a_at FOXN3 forkhead box N3 U68723.1 0.023025 11718869_x_at PALMD palmdelphin CF552454 0.023068 11727604_a_at EPB41L4A erythrocyte membrane protein NM_022140.3 0.023101 band 4.1 like 4A 11726633_s_at TRIM8 tripartite motif-containing 8 BCO21925.1 0.023187 11732370_a_at CUX1 cut-like homeobox 1 NM_181552.2 0.023258 11726113_a_at FAM46B family with sequence similarity NM_052943.3 0.023515 46, member B 11729100_a_at TTC18 tetratricopeptide repeat domain NM_145170.3 0.023862 18 11729259_a_at ZNF644 zinc finger protein 644 BQ014639 0.023971 11745806_a_at AMMECR1L AMME chromosomal region gene AK095871.1 0.024186 1-like 11717894_s_at PTP4AI protein tyrosine phosphatase BC023975.2 0.024306 type IVA, member 1 11728765_a_at PVRL4 poliovirus receptor-related 4 BC010423.1 0.024371 11740747_a_at DNMT3A DNA (cytosine-5-)- AF480163.1 0.024617 methyltransferase 3 alpha 11718868_a_at PALMD palmdelphin CF552454 0.025394 11758133_s_at COL4A3BP collagen, type IV, alpha 3 BE046819 0.026883 (Goodpasture antigen) binding protein 11749969_a_at TSPAN5 tetraspanin 5 AK295385.1 0.027108 11733899_a_at TROVE2 TROVE domain family, member 2 BX445026 0.027171 11747743_x_at MTF2 metal response element binding AK302776.1 0.027946 transcription factor 2 11746790_a_at BECN1 beclin 1, autophagy related AK298619.1 0.027999 11731573_a_at FRMD4B FERM domain containing 4B AU147415 0.028099 11724271_a_at HLF hepatic leukemia factor EL952952 0.028251 KRR1, small subunit (SSU) processome component, 11728683_x_at KRR1 homolog (yeast) U55766.1 0.02827 11758327_s_at BAZ1A bromodomain adjacent to zinc BF852255 0.02853 finger domain, 1A 11743300_a_at SRP72 signal recognition particle 72kDa AK225430.1 0.028582 11719103_at CPNE3 copine III CB250550 0.028667 11755895_a_at FAM129A family with sequence similarity AK095547.1 0.029014 129, member A 11721326_at C3orf14 chromosome 3 open reading AF236158.1 0.029423 frame 14 11738035_s_at RTN4 reticulon 4 AK302741.1 0.029458 11746122_s_at ZC3H11A zinc finger CCCH-type containing DA094705 0.029486 11A 11724085_at DAPL1 death associated protein-like 1 NM_001017920.2 0.030615 11735181_a_at DLX2 distal-less homeobox 2 NM_004405.3 0.030619

TABLE 18 Type III Epidermis; Down-regulated GeneTitan_ID Gene Title Public ID p 11743060_s_at COMMD10 COMM domain containing 10 AL572695 0.000211 11720515_s_at C9orf150 chromosome 9 open reading NM_203403.1 0.000414 frame 150 11716897_x_at PPIE peptidylprolyl isomerase E NM_006112.2 0.000535 (cyclophilin E) 11725787_a_at C4orf43 chromosome 4 open reading NM_018352.2 0.000571 frame 43 11729680_a_at KHK ketohexokinase (fructokinase) CR456801.1 0.000981 11745494_x_at ERCC8 excision repair cross- U28413.1 0.001042 complementing rodent repair deficiency, complementation group 8 11745948_a_at CHEK1 CHK1 checkpoint homolog (S. AK299783.1 0.0011 pombe) 11758965_at ATG4C ATG4 autophagy related 4 AK027773.1 0.001439 homolog C (S. cerevisiae) 11724079_s_at E2F2 E2F transcription factor 2 NM_004091.2 0.001667 11718280_s_at TRIAP1 TP53 regulated inhibitor of NM_016399.2 0.001683 apoptosis 1 11720459_s_at CAPRIN1 cell cycle associated protein 1 BQ002768 0.001735 11720615_a_at TUBG2 tubulin, gamma 2 NM_016437.2 0.00174 11757673_x_at RPL39 ribosomal protein L39 BX435916 0.001879 11720398_a_at NBN nibrin BC146797.1 0.002186 11734661_a_at CLSTN3 calsyntenin 3 NM_014718.3 0.0022 11726529_s_at BRCC3 BRCA1/BRCA2-containing NM_024332.2 0.002236 complex, subunit 3 11758291_s_at MRPS10 mitochondrial ribosomal BF701142 0.002439 protein S10 11726660_a_at GPN3 GPN-loop GTPase 3 AY359078.1 0.002577 11734619_x_at ALOX15B arachidonate 15- NM_001141.2 0.002749 lipoxygenase, type B 11719482_a_at MRPL21 mitochondrial ribosomal NM_181515.1 0.002874 protein L21 11716896_a_at PPIE peptidylprolyl isomerase E NM_006112.2 0.002936 (cyclophilin E) 11757498_s_at TMEM106C transmembrane protein 106C A1278554 0.003263 11722842_s_at ENAH enabled homolog (Drosophila) BC095481.1 0.003561 11722415_a_at HBS1L HBS1-like (S. cerevisiae) BC040849.1 0.003563 11742050_a_at API5 apoptosis inhibitor 5 AK294724.1 0.003612 11719499_at MAOB monoamine oxidase B NM_000898.4 0.003642 11717099_at HIST1H2BK histone cluster 1, H2bk NM_080593.1 0.003648 11744666_at FAN1 FANCD2/FANCI-associated BC047882.1 0.003687 nuclease 1 11756910_x_at FANCD2 Fanconi anemia, AL832427.1 0.00395 complementation group D2 11758535_s_at GPAM glycerol-3-phosphate AI074401 0.003969 acyltransferase, mitochondrial 11719920_at FXC1 fracture callus 1 homolog (rat) BC011014.1 0.004197 ring finger and CHY zinc finger 11719571_a_at RCHY1 domain containing 1 BC047393.1 0.004345 11763339_a_at SIVA1 SIVA1, apoptosis-inducing AK128704.1 0.004492 factor 11740706_a_at NFRKB nuclear factor related to NM_006165.3 0.004845 kappaB binding protein 11730410_a_at PXMP4 peroxisomal membrane NM_007238.4 0.005105 protein 4, 24 kDa 11739973_s_at NUAK1 NUAK family, SNF1-like BU686994 0.005564 kinase, 1 11757687_x_at DAD1 defender against cell death 1 BU535881 0.005572 11721430_a_at SYBU syntabulin (syntaxin- NM_001099744.1 0.005683 interacting) 11722555_sat HADH hydroxyacyl-CoA CR591982.1 0.005987 dehydrogenase 11736367_a_at MCM10 minichromosome NM_182751.1 0.006448 maintenance complex component 10 11739660_x_at PPCS phosphopantothenoylcysteine NM_001077447.1 0.0065 synthetase 11757604_a_at SAMM50 sorting and assembly BQ186212 0.006505 machinery component 50 homolog (S. cerevisiae) 11722398_s_at RWDD4 RWD domain containing 4 NM_152682.2 0.00654 11733866_a_at RARS2 arginyl-tRNA synthetase 2, NM_020320.3 0.006608 mitochondrial 11722351_at SRSF8 serine/arginine-rich splicing NM_032102.2 0.006693 factor 8 11726320_at ERO1L ERO1-like (S. cerevisiae) NM_014584.1 0.006774 up-regulated during skeletal muscle growth 5 homolog 11744384_x_at USMG5 (mouse) BC072683.1 0.006853 11733975_a_at DDHD2 DDHD domain containing 2 BU631346 0.007241 11727533_a_at FEZ2 fasciculation and elongation NM_001042548.1 0.007345 protein zeta 2 (zygin II) 11723462_a_at PHKB phosphorylase kinase, beta NM_001031835.2 0.007666 11718475_s_at IDH1 isocitrate dehydrogenase 1 NM_005896.2 0.007717 (NADP+), soluble 11744822_a_at NDUFB2 NADH dehydrogenase BC063026.1 0.007876 (ubiquinone) 1 beta subcomplex, 2, 8 kDa 11757589_a_at NDUFA12 NADH dehydrogenase BU537124 0.008475 (ubiquinone) 1 alpha subcomplex, 12 11726186_x_at C12orf48 chromosome 12 open reading NM_017915.2 0.008601 frame 48 11739972_at NUAK1 NUAK family, SNF1-like BU686994 0.008782 kinase, 1 11755294_x_at NEB nebulin BC063136.1 0.008985 11740962_a_at UBA5 ubiquitin-like modifier NM_198329.2 0.009092 activating enzyme 5 11753867_a_at NDUFA1 NADH dehydrogenase AB451304.1 0.009324 (ubiquinone) 1 alpha subcomplex, 1, 7.5 kDa 11757665_x_at NDUFS5 NADH dehydrogenase AA977996 0.009487 (ubiquinone) Fe-S protein 5, 15 kDa (NADH-coenzyme Q reductase) 11757684_a_at TPD52L2 tumor protein D52-like 2 A1806821 0.009563 11731068_s_at FIGNL1 fidgetin-like 1 NM_022116.3 0.009587 11743064_at CDC6 cell division cycle 6 homolog CR598029.1 0.009677 (S. cerevisiae) 11746036_s_at CBR1 carbonyl reductase 1 AK311219.1 0.009851 11729763_a_at LSM10 LSM10, U7 small nuclear RNA NM_032881.1 0.010114 associated 11719268_at TNNC1 troponin C type 1 (slow) NM_003280.2 0.010462 11758199_s_at RAD23B RAD23 homolog B BG571600 0.010584 (S. cerevisiae) 11723291_a_at NDUFA1 NADH dehydrogenase NM_004541.3 0.010596 (ubiquinone) 1 alpha subcomplex, 1, 7.5 kDa 11715771_a_at MRPL13 mitochondrial ribosomal NM_014078.4 0.010834 protein L13 11746174_s_at IDH1 isocitrate dehydrogenase 1 BC012846.1 0.011295 (NADP+), soluble 11724432_x_at TRAPPC2 trafficking protein particle NM_001011658.2 0.01144 complex 2 11746489_x_at GPAA1 glycosylphosphatidylinositol BC006383.2 0.011462 anchor attachment protein 1 homolog (yeast) 11724120_a_at TRIM59 tripartite motif-containing 59 NM_173084.2 0.01188 11764061_s_at NDUFB3 NADH dehydrogenase AA887183 0.012144 (ubiquinone) 1 beta subcomplex, 3, 12 kDa 11758083_s_at HPGD hydroxyprostaglandin AI743714 0.012171 dehydrogenase 15-(NAD) 11729715_a_at CBR1 carbonyl reductase 1 NM_001757.2 0.012312 11734864_x_at SARNP SAP domain containing NM_033082.3 0.012428 ribonucleoprotein 11717314_a_at HAUS1 HAUS augmin-like complex, NM_138443.3 0.012462 subunit 1 11751523_a_at TMED5 transmembrane emp24 AK293308.1 0.012569 protein transport domain containing 5 11754800_s_at GFM1 G elongation factor, AK092293.1 0.012699 mitochondrial 1 11746042_s_at TRA2B transformer 2 beta homolog AK098191.1 0.01285 (Drosophila) 11736741_a_at MKI67 antigen identified by NM_001145966.1 0.012861 monoclonal antibody Ki-67 11729333_at PADI1 peptidyl arginine deiminase, NM_013358.2 0.013091 type! 11751291_a_at SFXN4 sideroflexin 4 AY269785.1 0.013333 11717991_a_at SIDT2 SIDI transmembrane family, NM_001040455.1 0.013344 member 2 11748896_s_at CCRL1 chemokine (C-C motif) AK304461.1 0.013521 receptor-like 1 11716395_a_at GPR56 G protein-coupled receptor 56 NM_001145774.1 0.013559 11729716_s_at CBR1 carbonyl reductase 1 NM_001757.2 0.01399 11716063_at TNC tenascin C NM_002160.2 0.014267 11758011_x_at EEF1A1 eukaryotic translation BI495952 0.014362 elongation factor 1 alpha 1 11720317_a_at DAD1 defender against cell death 1 NM_001344.2 0.014754 11720186_s_at MAD2L1 MAD2 mitotic arrest deficient- NM_002358.3 0.014969 like 1 (yeast) 11725960_s_at CALM3 calmodulin 3 (phosphorylase NM_005184.2 0.015262 kinase, delta) 11730753_at AGPAT5 1-acylglycerol-3-phosphate 0- NM_018361.3 0.015361 acyltransferase 5 (lysophosphatidic acid acyltransferase, epsilon) 11735839_at STX19 syntaxin 19 NM_001001850.1 0.015426 11746655_a_at ACAA1 acetyl-CoA acyltransferase 1 AK303251.1 0.015895 11744002_s_at MTHFD2 methylenetetrahydrofolate BG026531 0.015966 dehydrogenase (NADP+ dependent) 2, methenyltetrahydrofolate cyclohydrolase 11721296_a_at NDUFB1 NADH dehydrogenase NM_004545.3 0.016187 (ubiquinone) 1 beta subcomplex, 1, 7 kDa 11725125_a_at NEB nebulin NM_004543.3 0.016335 11716624_s_at XPO1 exportin 1 (CRM1 homolog, NM_003400.3 0.016341 yeast) 11759922_a_at PARD3 par-3 partitioning defective 3 BC071566.1 0.016372 homolog (C. elegans)

TABLE 19 Type III Dermis; Up-regulated GeneTitan_ID Gene Title Public ID p 11733167_at LRRN4CL LRRN4 C-terminal like BC053902.1 0.000101 11716549_s_at ISLR immunoglobulin superfamily NM_005545.3 0.00021 containing leucine-rich repeat 11743191_a_at NTM neurotrimin AI343272 0.000486 11725753_a_at GRIA3 glutamate receptor, ionotrophic, U10301.1 0.000741 AMPA 3 11741377_a_at MMP2 matrix metallopeptidase 2 NM_001127891.1 0.001476 (gelatinase A, 72 kDa gelatinase, 72 kDa type IV collagenase) 11717765_a_at MGLL monoglyceride lipase NM_007283.5 0.001622 11721372_at TCF7L1 transcription factor 7-like 1 (T-cell NM_031283.1 0.001646 specific, HMG-box) 11722839_at LYAR Ly1 antibody reactive homolog AW958593 0.001955 (mouse) 11762135_at_ PTPRK protein tyrosine phosphatase, BC063596.1 0.002158 receptor type, K 11721467_s_at CD276 CD276 molecule NM_001024736.1 0.002325 11761134_at MYST3 MYST histone acetyltransferase BC142959.1 0.003233 (monocytic leukemia) 3 11720440_at OLFML2B olfactomedin-like 2B NM_015441.1 0.003454 11745431_a_at SVIL supervillin BC092440.1 0.003484 11739746_s_at SVIL supervillin CD366976 0.003636 11757808_s_at RERE arginine-glutamic acid dipeptide BM706668 0.003739 (RE) repeats 11725937_a_at LGALS3 lectin, galactoside-binding, BC053667.1 0.003825 soluble, 3 11720274_x_at ALKBH6 alkB, alkylation repair homolog 6 NM_032878.3 0.003968 (E. coli) 11755955_a_at FAP fibroblast activation protein, alpha AL832166.1 0.003989 11724619_at RSPO3 R-spondin 3 homolog (Xenopus NM_032784.3 0.004121 laevis) 11729170_x_at DUSP10 dual specificity phosphatase 10 AF179212.1 0.004193 11752038_a_at AQPEP laeverin BC068560.1 0.004865 11720846_at CD248 CD248 molecule, endosialin NM_020404.2 0.005206 11731143_a_at GPR133 G protein-coupled receptor 133 NM_198827.3 0.005394 11728451_a_at PCOLCE2 procollagen C-endopeptidase NM_013363.2 0.005523 enhancer 2 11731682_at CD70 CD70 molecule NM_001252.3 0.005777 11716226_a_at LIMA1 LIM domain and actin binding 1 BC136763.1 0.006288 11750244_a_at MGLL monoglyceride lipase AK304844.1 0.00649 11762370_x_at BNC1 basonuclin 1 L03427.1 0.006531 11729101_a_at AKR1C2 aldo-keto reductase family 1, NM_205845.1 0.006587 member C2 (dihyd rod iol dehydrogenase 2; bile acid binding protein; 3-alpha hydroxysteroid dehydrogenase, type III) 11731649_x_at NTM neurotrimin AY358331.1 0.006934 11716238_at ARHGAP1 Rho GTPase activating protein 1 NM_004308.2 0.006953 11728498_a_at SVIL supervillin NM_003174.3 0.00696 11725517_x_at ABCG1 ATP-binding cassette, sub-family G NM_207627.1 0.007347 (WHITE), member 1 11728605_s_at LIMS1 LIM and senescent cell antigen- NM_033514.2 0.007353 like domains 1 11752843_x_at SQSTM1 sequestosome 1 AK304877.1 0.007432 11757557_s_at CADM1 cell adhesion molecule 1 H23245 0.007475 11718269_x_at ANGPTL2 angiopoietin-like 2 AY358274.1 0.007782 11747944_a_at PPFIA2 protein tyrosine phosphatase, AK296380.1 0.008378 receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 2 11761149_a_at C5orf45 chromosome 5 open reading AK293901.1 0.008409 frame 45 11737357_a_at CNGA3 cyclic nucleotide gated channel NM_001298.2 0.008783 alpha 3 11743250_a_at MMP2 matrix metallopeptidase 2 BX357054 0.009096 (gelatinase A, 72 kDa gelatinase, 72 kDa type IV collagenase) 11725515_a_at ABCG1 ATP-binding cassette, sub-family G NM_207627.1 0.009131 (WHITE), member 1 11759362_x_at PHKG1 phosphorylase kinase, gamma 1 BC051327.1 0.009194 (muscle) 11717802_s_at ATF5 activating transcription factor 5 BE300055 0.009354 11723070_a_at CYTL1 cytokine-like 1 NM_018659.2 0.009527 11731650_a_at NTM neurotrimin NM_001048209.1 0.009599 11720845_a_at CD248 CD248 molecule, endosialin NM_020404.2 0.009838 11716322_s_at PRKCDBP protein kinase C, delta binding NM_145040.2 0.009887 protein 11718658_s_at CD34 CD34 molecule NM_001773.2 0.010659 11747945_x_at PPFIA2 protein tyrosine phosphatase, AK296380.1 0.010696 receptor type, f polypeptide (PTPRF), interacting protein (liprin), alpha 2 11717764_x_at MGLL monoglyceride lipase BC006230.2 0.010744 11743251_s_at MMP2 matrix metallopeptidase 2 BX357054 0.010854 (gelatinase A, 72 kDa gelatinase, 72 kDa type IV collagenase) 11731303_a_at DUSP10 dual specificity phosphatase 10 BC020608.1 0.010964 11719737_a_at FAM134B family with sequence similarity BC053326.1 0.011265 134, member B 11758143_s_at DUSP8 dual specificity phosphatase 8 BE350906 0.011421 11724441_x_at PTGIS prostaglandin 12 (prostacyclin) NM_000961.3 0.011582 synthase 11737583_s_at SGCD sarcoglycan, delta (35 kDa NM_001128209.1 0.01169 dystrophin-associated glycoprotein) 11729541_a_at CAMKK2 calcium/calmodulin-dependent AB081337.1 0.011881 protein kinase kinase 2, beta 11737108_a_at CCRL1 chemokine (C-C motif) receptor- NM_178445.1 0.012 like 1 11721507_at DVL3 dishevelled, dsh homolog 3 NM_004423.3 0.012974 (Drosophila) 11750245_x_at MGLL monoglyceride lipase AK304844.1 0.013107 11737946_a_at XPNPEP2 X-prolyl aminopeptidase BC143901.1 0.01347 (aminopeptidase P) 2, membrane- bound 11727155_a_at TRIOBP TRIO and F-actin binding protein NM_007032.5 0.013526 11720441_x_at OLFML2B olfactomedin-like 2B NM015441.1 0.01374 11727773_at LARP6 La ribonucleoprotein domain NM_197958.1 0.013995 family, member 6 11728499_x_at SVIL supervillin NM_003174.3 0.01402 11745659_s_at POM121 POM121 membrane glycoprotein BC130587.1 0.014097 11752562_x_at CDH13 cadherin 13, H-cadherin (heart) AK294277.1 0.014197 11720617_at TRIM9 tripartite motif-containing 9 NM_015163.5 0.014562 11757548_s_at ADAMTSL1 ADAMTS-like 1 DB329733 0.014859 11753179_s_at FAM134B family with sequence similarity BC030517.1 0.014991 134, member B 11729285_a_at NFU1 NFU1 iron-sulfur cluster scaffold homolog (S. cerevisiae) NM_001002755.1 0.01519 11741286_a_at CCRL1 chemokine (C-C motif) receptor- AF110640.1 0.015787 like 1 11732315_a_at SGCD sarcoglycan, delta (35 kDa AF010236.1 0.015795 dystrophin-associated glycoprotein) 11715852_at PDGFRB platelet-derived growth factor NM_002609.3 0.016136 receptor, beta polypeptide 11730404_at MEX3B mex-3 homolog B (C. elegans) NM_032246.3 0.0163 11751986_at MMP19 matrix metallopeptidase 19 U38320.1 0.016486 11731122_a_at VASH2 vasohibin 2 BC051856.1 0.016505 11732785_a_at C16orf45 chromosome 16 open reading NM_001142469.1 0.017241 frame 45 11757765_s_at SGCD sarcoglycan, delta (35 kDa AA401248 0.017347 dystroph in-associated glycoprotein) 11743143_at COX11 COX11 cytochrome c oxidase AK293851.1 0.017504 assembly homolog (yeast) 11724142_s_at RAB11FIP2 RAB11 family interacting protein 2 DB356544 0.017868 (class I) 11723075_a_at BCL9L B-cell CLL/lymphoma 9-like AY296059.1 0.017989 11747704_a_at CLDN11 claudin 11 AK294087.1 0.017998 11716376_at SERPINA5 serpin peptidase inhibitor, clade A NM_000624.4 0.018046 (alpha-1 antiproteinase, antitrypsin), member 5 11756879_a_at STARD9 StAR-related lipid transfer (START) CR936665.1 0.018874 domain containing 9 11733166_at LRRN4CL LRRN4 C-terminal like NM_203422.1 0.018933 11720163_at VEGFC vascular endothelial growth factor NM_005429.2 0.018951 C 11754821_s_at SLC38A1 solute carrier family 38, member 1 AI476037 0.019062 11720082_at CBX6 chromobox homolog 6 NM_014292.3 0.020169 11762231_x_at BBS1 Bardet-Biedl syndrome 1 AK294962.1 0.020213 11732462_at ADAMTSL1 ADAMTS-like 1 AK123028.1 0.020317 11761563_x_at HEATR1 HEAT repeat containing 1 BC062442.1 0.0204 11727714_at KCNJ12 potassium inwardly-rectifying NM_021012.4 0.020553 channel, subfamily J, member 12 11727780_a_at SCARA5 scavenger receptor class A, NM_173833.4 0.020636 member 5 (putative) 11749436_a_at NFIC nuclear factor I/C (CCAAT-binding AK297825.1 0.020877 transcription factor) 11731209_s_at C15orf59 chromosome 15 open reading NM_001039614.1 0.021422 frame 59 11727125_a_at PVRL3 poliovirus receptor-related 3 BE544927 0.021561 11744741_at LOH3CR2A loss of heterozygosity, 3, AF086709.2 0.021592 chromosomal region 2, gene A 11717891_a_at ECM1 extracellular matrix protein 1 BC023505.2 0.021868

TABLE 20 Type III Dermis; Down-regulated GeneTitan_ID Gene Title Public ID p 11727158_a_at STRBP spermatid perinuclear NM_018387.3 0.000131 RNA binding protein 11756850_x_at CCT8 chaperonin containing CR612497.1 0.000172 TCP1, subunit 8 (theta) 11754000_x_at CD58 CD58 molecule CR456939.1 0.000222 11737761_a_at HSD17B4 hydroxysteroid (17-beta) dehydrogenase 4 NM_000414.2 0.000267 11754276_a_at RAD23B RAD23 homolog B (S. BG501496 0.000343 cerevisiae) 11743094_at SPRR4 small proline-rich protein BC069445.1 0.000362 4 11724156_at SULT1E1 sulfotransferase family NM_005420.2 0.000395 1E, estrogen-preferring, member 1 11749267_a_at SRD5A1 steroid-5-alpha- AK315996.1 0.000399 reductase, alpha polypeptide 1 (3-oxo-5 alpha-steroid delta 4- dehydrogenase alpha 1) 11720183_s_at EEF1B2 eukaryotic translation NM_001959.3 0.000428 elongation factor 1 beta 2 11737053_s_at HSPD1 heat shock 60 kDa protein NM_002156.4 0.000467 1 (chaperonin) 11740377_a_at PXMP4 peroxisomal membrane AK297018.1 0.000581 protein 4, 24 kDa 11726318_s_at EEF1G eukaryotic translation NM_001404.4 0.000583 elongation factor 1 gamma 11720184_x_at EEF1B2 eukaryotic translation NM_001959.3 0.000595 elongation factor 1 beta 2 11746149_x_at BCHE butyrylcholinesterase M16474.1 0.000611 11737762_x_at HSD17B4 hydroxysteroid (17-beta) NM_000414.2 0.000646 dehydrogenase 4 11739725_a_at TC2N tandem C2 domains, NM_001128595.1 0.000648 nuclear 11754918_s_at HMGCS1 3-hydroxy-3- AK095492.1 0.000648 methylglutaryl-CoA synthase 1 (soluble) 11741799_a_at BCOR BCL6 corepressor AF317391.1 0.000649 11736831_a_at SEC23B Sec23 homolog B (S. NM_032986.3 0.000656 cerevisiae) 11744777_s_at DPY30 dpy-30 homolog (C. BC015970.1 0.000676 elegans) 11754418_s_at G3BP1 GTPase activating protein AK130003.1 0.000682 (SH3 domain) binding protein 1 11723250_a_at EML2 echinoderm microtubule NM_012155.1 0.00072 associated protein like 2 11752369_a_at IMPDH2 IMP (inosine 5′- AK293397.1 0.000726 monophosphate) dehydrogenase 2 11729643_s_at TPD52 tumor protein D52 CB219128 0.000732 11755057_s_at ATP2C1 ATPase, Ca++ AB037768.1 0.000739 transporting, type 2C, member 1 11716946_s_at TM9SF3 transmembrane 9 AF269150.1 0.000773 superfamily member 3 11756300_a_at ANP326 acidic (leucine-rich) BX432546 0.000781 nuclear phosphoprotein 32 family, member B 11716134_a_at MTOR mechanistic target of NM_004958.3 0.000785 rapamycin (serine/threonine kinase) 11755203_x_at RPL21 ribosomal protein L21 BX647669.1 0.000869 11730938_x_at PYCR1 pyrroline-5-carboxylate NM_153824.1 0.000877 reductase 1 11750545_a_at CNOT7 CCR4-NOT transcription BC007315.2 0.000884 complex, subunit 7 11727826_a_at C2orf56 chromosome 2 open BC004548.2 0.00093 reading frame 56 11718344_a_at CNOT7 CCR4-NOT transcription NM_013354.5 0.000947 complex, subunit 7 11756600_a_at TPD52 tumor protein D52 AK308983.1 0.000999 11734619_x_at ALOX156 arachidonate 15- NM_001141.2 0.001036 lipoxygenase, type B 11715621_at UFC1 ubiquitin-fold modifier NM_016406.3 0.001145 conjugating enzyme 1 11715958_s_at RPL7 ribosomal protein L7 NM_000971.3 0.001233 11748713_a_at ASPM asp (abnormal spindle) AY971957.1 0.001256 homolog, microcephaly associated (Drosophila) 11758707_s_at C5orf25 chromosome 5 open DB526316 0.001417 reading frame 25 200081_PM_s_at RPS6 ribosomal protein S6 BE741754 0.00144 11726299_x_at LGALS8 lectin, galactoside- AF342815.1 0.001452 binding, soluble, 8 11756210_a_at RCL1 RNA terminal phosphate AL582781 0.001477 cyclase-like 1 11743604_s_at RRM1 ribonucleotide reductase BE618815 0.001487 M1 11729641_a_at TPD52 tumor protein D52 BG389015 0.001493 11718461_at SLC39A11 solute carrier family 39 NM_139177.3 0.001532 (metal ion transporter), member 11 11725053_x_at TOP1MT topoisomerase (DNA) I, NM_052963.1 0.001534 mitochondrial 11758027_s_at HOOK1 hook homolog 1 CD243255 0.001606 (Drosophila) 11745205_s_at TPD52 tumor protein D52 BC018117.1 0.001623 11760342_a_at PPP3CB protein phosphatase 3, M29550.1 0.001681 catalytic subunit, beta isozyme 11725875_at WDR66 WD repeat domain 66 NM_144668.4 0.001791 11739308_s_at DLG1 discs, large homolog 1 BM681931 0.001815 (Drosophila) 11719666_a_at STMN1 stathmin 1 BC082228.1 0.001852 11752283_a_at ALOX15B arachidonate 15- AK298095.1 0.001904 lipoxygenase, type B 11723312_a_at PXMP2 peroxisomal membrane NM_018663.1 0.001916 protein 2, 22 kDa 11719667_s_at STMN1 stathmin 1 BC082228.1 0.001961 11728791_at THRSP thyroid hormone NM_003251.2 0.001966 responsive 11734917_a_at METTL4 methyltransferase like 4 BQ009802 0.001983 11717236_x_at RPS7 ribosomal protein S7 NM_001011.3 0.002022 11754132_x_at COMT catechol-0- BT007125.1 0.002101 methyltransferase 11743372_s_at PTGES3 prostaglandin E synthase CR611609.1 0.002174 3 (cytosolic) 11730411_a_at PXMP4 peroxisomal membrane BF057649 0.002209 protein 4, 24 kDa 200063_PM_s_at NPM1 nucleophosmin BC002398.1 0.002233 (nucleolar phosphoprotein B23, numatrin) 11722642_a_at DGAT2 diacylglycerol 0- BC015234.1 0.002305 acyltransferase 2 11752550_x_at CCT8 chaperonin containing AK293705.1 0.002336 TCP1, subunit 8 (theta) 11758217_s_at FAM108C1 family with sequence CB997200 0.002358 similarity 108, member C1 11717182_a_at PDS5A PDS5, regulator of NM_001100399.1 0.002387 cohesion maintenance, homolog A (S. cerevisiae) 11717153_a_at C20orf3 chromosome 20 open NM_020531.2 0.002394 reading frame 3 11742779_a_at HIBCH 3-hydroxyisobutyryl-CoA U66669.1 0.002431 hydrolase 11744264_a_at SEC11C SEC11 homolog C (S. AI816180 0.002433 cerevisiae) 11753788_x_at CDKN3 cyclin-dependent kinase AF213040.1 0.002434 inhibitor 3 11758709_s_at RDH11 retinol dehydrogenase 11 A1972157 0.002449 (all-trans/9-cis/11-cis) 11727320_at IGFL2 IGF-like family member 2 NM_001002915.2 0.002476 11730803_a_at PRPF38B PRP38 pre-mRNA NM_018061.2 0.002515 processing factor 38 (yeast) domain containing B 11753740_x_at CYB5A cytochrome b5 type A CR456990.1 0.002572 (microsomal) 11718246_a_at KIAA0146 KIAA0146 NM_001080394.1 0.002609 11720768_at METTL9 methyltransferase like 9 NM_016025.3 0.002613 11755017_a_at CHCHD7 coiled-coil-helix-coiled- AK098285.1 0.002843 coil-helix domain containing 7 11732128_s_at CCT4 chaperonin containing BC106934.1 0.002855 TCP1, subunit 4 (delta) 11744900_x_at FADS2 fatty acid desaturase 2 AF108658.1 0.002865 11715881_a_at DAP3 death associated protein NM_004632.2 0.002925 3 11756875_x_at COMMD6 COMM domain CR603325.1 0.002962 containing 6 11756783_a_at TF transferrin BC045772.1 0.002967 11723197_at HNRNPA3 heterogeneous nuclear BX434302 0.003022 ribonucleoprotein A3 11729941_at TMEM56 transmembrane protein NM_152487.2 0.003035 56 11716813_a_at GATM glycine AK298350.1 0.003053 amidinotransferase (L- arginine:glycine amidinotransferase) 11721242_s_at FDFT1 farnesyl-diphosphate NM_004462.3 0.003125 farnesyltransferase 1 11749786_x_at HNRNPF heterogeneous nuclear AK296696.1 0.003153 ribonucleoprotein F 11723313_s_at PXMP2 peroxisomal membrane NM_018663.1 0.003162 protein 2, 22 kDa 11727286_a_at ZNF323 zinc finger protein 323 NM_001135215.1 0.003185 11720813_at INTS10 integrator complex NM_018142.2 0.003216 subunit 10 11749874_a_at OXCT1 3-oxoacid CoA AK299668.1 0.003244 transferase 1 11757320_x_at CYB5A cytochrome b5 type A AA706740 0.003263 (microsomal) 11733591_a_at C1orf204 chromosome 1 open NM_001134233.1 0.003297 reading frame 204 11718135_at PRPS2 phosphoribosyl NM_001039091.1 0.003316 pyrophosphate synthetase 2 11716302_s_at ACSL1 acyl-CoA synthetase long- NM_001995.2 0.003359 chain family member 1 11744392_a_at PAPOLA poly(A) polymerase alpha BC000927.1 0.003388 11741012_a_at SC4MOL sterol-C4-methyl oxidase- AK292418.1 0.00342 like synovial sarcoma translocation, 11722800_a_at SS18 chromosome 18 CB241009 0.003458 11755439_x_at UBAC2 UBA domain containing 2 BC053346.1 0.003608 11756674_s_at STRBP spermatid perinuclear CR596677.1 0.003641 RNA binding protein

The gene expression signatures provided in Table 9-20 can be used to classify the type of periorbital dyschromia exhibited by person. For example, a person can be classified as exhibiting Type I periorbital dyschromia when the gene expression signature of a skin tissue sample obtained from the periorbital region of the person corresponds to 15% or more (e.g., more than 25%, 50%, 75%, 80%, 90% or even up to 100%) of the genes from Tables 9 to 12. In other words, a person can be classified as exhibiting Type I periorbital dyschromia if the top 100 upregulated genes in the epidermis comprise 15% or more the genes in Table 9; the top 100 downregulated genes in the epidermis comprise 15% or more of the genes in Table 10; the top 100 upregulated genes in the dermis comprise 15% or more of the genes in Table 11; and/or the top 100 downregulated genes in the dermis comprise 15% or more of the genes in Table 12. Similarly, a person can be classified as exhibiting Type II or Type III periorbital dyschromia when the gene expression signature of the skin tissue sample comprises significantly upregulated and downregulated genes that correspond to 15% or more of the genes in at least one of Tables 13 to 16 or Tables 17 to 20, respectively.

Theme analysis may be used to identify biological or phenotypic themes associated with the gene expression data that correspond to Type I, Type II and Type III periorbital dyschromia. Theme analysis is a statistical analysis-based method for detecting biological patterns in gene expression data. The method uses an ontology of controlled vocabulary terms developed by the Gene Ontology (“GO”) Consortium [Ashburner, M. et al. (2000) Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet, 25, 25-29] that describe the biological processes, molecular functions and cellular components associated with gene products. Analysis involves statistical comparison of a regulated list of genes and a larger reference list of all the expressed genes, to determine if genes annotated to specific GO terms are significantly enriched in the regulated list. This analysis may reveal biological patterns when multiple genes associated with a given GO term occur on the regulated list at a frequency greater than expected by chance. Such analysis may be performed using Theme Extractor proprietary software and an algorithm that calculates the p-value of each ontology term. Data may be analyzed for statistical significance, for example, by the Fisher's exact test. Conventional approaches and statistical methods such as, for example, Gene Set Enrichment Analysis described by Subramanian, A., et al., in “Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles,” Proc. Natl. Acad Sci U.S.A, 102, 15545-155501 (2005) are suitable for conducting theme analysis herein.

Table 21 shows Gene Ontology Biological Process terms that are significantly enriched in the epidermis of subjects with Type I periorbital dyschromia. The results in Table 21 were generated from differentially expressed genes in those subjects with top ranked genes shown in Tables 13 and 14. Only the most highly significant themes are shown (p-value≤1×10⁻² to p≥1×10⁻⁵) and the theme analysis was done separately for the up- and down-regulated genes. The level of indentation in the terms column (i.e., the number of dots preceding the term) generally indicates the level in the GO hierarchy and parent/child relationships between terms.

TABLE 21 Type I Epidermis vs N-D Gene Ontology Biological Process Terms Up Down Directional GO:0002764 immune response-regulating ** signaling pathway GO:0002758 innate immune response-activating *** signal transduction GO:0002224 toll-like receptor signaling pathway ** GO:0048468 cell development ** GO:0002376 immune system process **** GO:0002252 immune effector process **** GO:0002253 activation of immune response **** GO:0002218 activation of innate immune *** response GO:0006955 immune response **** GO:0045087 innate immune response **** GO:0071383 cellular response to steroid ** hormone stimulus GO:0006954 inflammatory response * GO:0045047 protein targeting to ER **** GO:0032984 macromolecular complex **** disassembly GO:0016568 chromatin modification * GO:0050776 regulation of immune response **** GO:0006749 glutathione metabolic process ** GO:0019752 carboxylic acid metabolic process * GO:0007154 cell communication * GO:0016042 lipid catabolic process * GO:0070588 calcium ion transmembrane * transport *p-value between 1 × 10⁻² and 1 × 10⁻³ **p-value between 1 × 10⁻³ and 1 × 10⁻⁴ ***p-value between 1 × 10⁻⁴ and 1 × 10⁻⁵ ****p-value less than 1 × 10⁻⁵

Table 21 shows Gene Ontology Biological Process terms that are significantly enriched in the epidermis of subjects with Type II periorbital dyschromia. The results in Table 14 were generated from differentially expressed genes in those subjects with top ranked genes shown in Tables 13 and 14. Only the most highly significant themes are shown (p≤1×10⁻²), and the theme analysis was done separately for the up- and down-regulated genes.

TABLE 22 Terms Type II Epidermis vs N-D Gene Ontology Biological Process Up Down Directional GO:0042770 signal transduction in **** response to DNA damage GO:0072331 signal transduction by **** p53 class mediator GO:0007173 epidermal growth factor ** receptor signaling pathway GO:0001942 hair follicle development **** GO:0008544 epidermis development **** GO:0045444 fat cell differentiation * GO:0019882 antigen processing and **** presentation GO:0071453 cellular response to oxygen ** levels GO:0071456 cellular response to hypoxia ** GO:0031960 response to corticosteroid * stimulus GO:0033554 cellular response to stress **** GO:0006281 DNA repair ** GO:0000077 DNA damage checkpoint **** GO:0006950 response to stress ** GO:0009268 response to pH ** GO:0006886 intracellular protein **** transport GO:0006605 protein targeting **** GO:0015031 protein transport **** GO:0046907 intracellular transport **** GO:0048193 Golgi vesicle transport ** GO:0006839 mitochondrial transport **** GO:0015986 ATP synthesis coupled *** proton transport GO:0016568 chromatin modification ** GO:0031326 regulation of cellular ** biosynthetic process GO:0006521 regulation of cellular amino **** acid metabolic process GO:0010565 regulation of cellular ketone **** metabolic process GO:0051171 regulation of nitrogen compound metabolic process GO:0051726 regulation of cell cycle ** GO:0045454 cell redox homeostasis ** GO:0032652 regulation of interleukin-1 ** production GO:0035383 thioester metabolic process **** GO:0006637 acyl-CoA metabolic process **** GO:0006099 tricarboxylic acid cycle **** GO:0006635 fatty acid beta-oxidation ** GO:0018108 peptidyl-tyrosine ** phosphorylation GO:0006082 organic acid metabolic **** process GO:0006520 cellular amino acid ** metabolic process GO:0043038 amino acid activation **** GO:0006091 generation of precursor **** metabolites and energy GO:0006096 glycolysis ** GO:0006119 oxidative phosphorylation **** GO:0042773 ATP synthesis coupled **** electron transport GO:0045333 cellular respiration **** GO:0007049 cell cycle **** GO:0005975 carbohydrate metabolic *** process GO:0006007 glucose catabolic process *** GO:0016052 carbohydrate catabolic **** process GO:0055114 oxidation-reduction process ****

Table 23 shows Gene Ontology Biological Process terms that are significantly enriched in the epidermis of subjects with Type III periorbital dyschromia. The results in Table 23 were generated differentially expressed genes in those subjects with top ranked genes shown in Tables 17 and 18. Only the most highly significant themes are shown (p-value <1×10⁻²), and the theme analysis was done separately for the up- and down-regulated genes.

TABLE 23 Type III Epidermis vs N-D Gene Ontology Biological Process Terms Up Down Directional GO:0048731 system development ** GO:0048856 anatomical structure development *** GO:0030154 cell differentiation ** GO:0070482 response to oxygen levels * GO:0033365 protein localization to organelle ** GO:0045047 protein targeting to ER ** GO:0006605 protein targeting ** GO:0006612 protein targeting to membrane ** GO:0006839 mitochondrial transport ** GO:0001934 positive regulation of protein ** phosphorylation GO:0051347 positive regulation of transferase ** activity GO:0010627 regulation of intracellular protein ** kinase cascade GO:0080135 regulation of cellular response * to stress GO:0051320 S phase *** GO:0006414 translational elongation **** GO:0006091 generation of precursor *** metabolites and energy GO:0006119 oxidative phosphorylation ** GO:0022900 electron transport chain **** GO:0045333 cellular respiration **** GO:0055114 oxidation-reduction process ***

Table 24 shows Gene Ontology Biological Process terms that are significantly enriched in the dermis of subjects with Type I periorbital dyschromia. The results in Table 24 were generated from differentially expressed genes in those subjects with top ranked genes shown in Tables 11 and 12. Only the most highly significant themes are shown (p≤1×10⁻²), and the theme analysis was done separately for the up- and down-regulated genes.

TABLE 24 Type I Dermis vs N-D Gene Ontology Biological Process Terms Up Down Directional GO:0000165 MAPK cascade ** GO:0048011 nerve growth factor receptor * signaling pathway GO:0007179 transforming growth factor beta * receptor signaling pathway GO:0050877 neurological system process *** GO:0007596 blood coagulation * GO:0050878 regulation of body fluid levels * GO:0001944 vasculature development **** GO:0001568 blood vessel development **** GO:0048514 blood vessel morphogenesis **** GO:0001525 angiogenesis **** GO:0030183 B cell differentiation * GO:0007399 nervous system development **** GO:0022008 neurogenesis *** GO:0048699 generation of neurons *** GO:0030182 neuron differentiation ** GO:0048666 neuron development ** GO:0048870 cell motility ** GO:0043627 response to estrogen stimulus ** GO:0006281 DNA repair **** GO:0001666 response to hypoxia * GO:0006979 response to oxidative stress * GO:0009611 response to wounding *** GO:0070482 response to oxygen levels ** GO:0033365 protein localization to organelle **** GO:0045047 protein targeting to ER **** GO:0006605 protein targeting **** GO:0015031 protein transport **** GO:0046907 intracellular transport **** GO:0034330 cell junction organization ** GO:0030198 extracellular matrix organization * GO:0007010 cytoskeleton organization * GO:0051276 chromosome organization *** GO:0032200 telomere organization ** GO:0006338 chromatin remodeling * GO:0043408 regulation of MAPK cascade *** GO:0051924 regulation of calcium ion transport * GO:0010827 regulation of glucose transport *** GO:0030334 regulation of cell migration **** GO:0010564 regulation of cell cycle process ** GO:0010646 regulation of cell communication **** GO:0022402 cell cycle process ** GO:0022403 cell cycle phase *** GO:0007049 cell cycle ** GO:0000278 mitotic cell cycle **** GO:0007154 cell communication **** GO:0008202 steroid metabolic process * GO:0006694 steroid biosynthetic process ** GO:0016126 sterol biosynthetic process **** GO:0006695 cholesterol biosynthetic process **** GO:0061061 muscle structure development *

Table 25 shows Gene Ontology Biological Process terms that are significantly enriched in the dermis of subjects with Type II periorbital dyschromia. The results in Table 25 were generated from differentially expressed genes in those subjects with top ranked genes shown in Tables 15 and 16. Only the most highly significant themes are shown (p<1×10⁻²), and the theme analysis was done separately for the up- and down-regulated genes.

TABLE 25 Type II Dermis vs. N-D Gene Ontology Biological Process Terms Up Down Directional GO:0019226 transmission of nerve impulse * GO:0050877 neurological system process ** GO:0007399 nervous system development ** GO:0022008 neurogenesis **** GO:0048699 generation of neurons *** GO:0030182 neuron differentiation *** GO:0048666 neuron development *** GO:0031175 neuron projection development ** GO:0007409 axonogenesis ** GO:0048667 cell morphogenesis involved in ** neuron differentiation GO:0048468 cell development *** GO:0006281 DNA repair **** GO:0033365 protein localization to organelle **** GO:0070972 protein localization to endoplasmic **** reticulum GO:0006605 protein targeting **** GO:0006612 protein targeting to membrane **** GO:0015031 protein transport **** GO:0006810 transport **** GO:0051641 cellular localization **** GO:0016043 cellular component organization **** GO:0022411 cellular component disassembly **** GO:0030198 extracellular matrix organization * GO:0032984 macromolecular complex **** disassembly GO:0043624 cellular protein complex **** disassembly GO:0071156 regulation of cell cycle arrest ** GO:0010646 regulation of cell communication * GO:0051320 S phase ** GO:0044248 cellular catabolic process **** GO:0006457 protein folding ** GO:0007049 cell cycle * GO:0000278 mitotic cell cycle ** GO:0007154 cell communication * GO:0006694 steroid biosynthetic process ** GO:0006695 cholesterol biosynthetic process ***

Table 26 shows Gene Ontology Biological Process terms that are significantly enriched in the epidermis of subjects with Type III periorbital dyschromia. The results in Table 26 were generated from differentially expressed genes in those subjects with top ranked genes shown in Tables 19 and 20. Only the most highly significant themes are shown (p≤1×10⁻²), and the theme analysis was done separately for the up- and down-regulated genes.

TABLE 26 Type III Dermis vs. N-D Gene Ontology Biological Process Terms Up Down Directional GO:0007155 cell adhesion ** GO:0019226 transmission of nerve impulse * GO:0051403 stress-activated MAPK cascade *** GO:0007254 JNK cascade ** GO:0000165 MAPK cascade **** GO:0016055 Wnt receptor signaling pathway * GO:0050877 neurological system process ** GO:0001944 vasculature development *** GO:0001568 blood vessel development ** GO:0048514 blood vessel morphogenesis ** GO:0022008 neurogenesis ** GO:0030182 neuron differentiation ** GO:0048666 neuron development ** GO:0061061 muscle structure development ** GO:0042692 muscle cell differentiation ** GO:0048646 anatomical structure formation **** involved in morphogenesis GO:0033554 cellular response to stress * GO:0006281 DNA repair **** GO:0001666 response to hypoxia ** GO:0070482 response to oxygen levels *** GO:0036293 response to decreased oxygen levels ** GO:0034613 cellular protein localization **** GO:0033365 protein localization to organelle **** GO:0070972 protein localization to endoplasmic **** reticulum GO:0045047 protein targeting to ER **** GO:0006886 intracellular protein transport **** GO:0006605 protein targeting **** GO:0006612 protein targeting to membrane **** GO:0015031 protein transport **** GO:0006811 ion transport *** GO:0006812 cation transport ** GO:0030001 metal ion transport ** GO:0050000 chromosome localization **** GO:0022411 cellular component disassembly **** GO:0034330 cell junction organization * GO:0034622 cellular macromolecular complex **** assembly GO:0007010 cytoskeleton organization ** GO:0000226 microtubule cytoskeleton ** organization GO:0070925 organelle assembly **** GO:0051276 chromosome organization *** GO:0032200 telomere organization *** GO:0000819 sister chromatid segregation **** GO:0043408 regulation of MAPK cascade **** GO:0010646 regulation of cell communication **** GO:0019725 cellular homeostasis * GO:0022402 cell cycle process **** GO:0035383 thioester metabolic process ** GO:0071616 acyl-CoA biosynthetic process ** GO:0035337 fatty-acyl-CoA metabolic process *** GO:0044248 cellular catabolic process **** GO:0046394 carboxylic acid biosynthetic process *** GO:0044255 cellular lipid metabolic process * GO:0006631 fatty acid metabolic process ** GO:0019752 carboxylic acid metabolic process **** GO:0007049 cell cycle **** GO:0007154 cell communication *** GO:0005975 carbohydrate metabolic process ** GO:0006694 steroid biosynthetic process ** GO:0006695 cholesterol biosynthetic process **** GO:0008610 lipid biosynthetic process ** GO:0055114 oxidation-reduction process **

The theme analysis of Type I periorbital dyschromia may be broadly correlated to cellular physiological responses associated with cell stress and cell communication in the epidermis and metabolism, stress and structure in the dermis. In particular, a theme analysis of epidermal gene expression related to Type I periorbital dyschromia revealed, generally, an increase in gene expression associated with calcium ion trans-membrane transport, chromatin modification and cell communication and a decrease in gene expression associated with immune response and toll-like receptor pathway. Dermal gene expression revealed the following general themes: an increase in gene expression associated with response to estrogen stimulus, regulation of body fluid levels, regulation of cell migration, hypoxic stress, oxidative damage, response to wounding, blood vessel development, extracellular matrix organization, cytoskeleton organization, cell junction organization, neuron development and muscle structure development and a decrease in gene expression associated with cholesterol synthesis, regulation of glucose transport and protein transport.

The theme analysis of Type II periorbital dyschromia appears to show a general correlation with the cellular physiological responses associated with metabolism and energy production, cell stress and cell communication in the epidermis and metabolism and structure in the dermis. In particular, a theme analysis of epidermal gene expression related to Type II periorbital dyschromia revealed, generally, an increase in gene expression associated with chromatin modification and a decrease in gene expression associated with glycolysis, ATP synthesis, cell cycle, TCA cycle, response to pH, antigen presentation and processing. Dermal gene expression revealed the following general themes: an increase in gene expression associated with transmission of nerve impulses and neuron development and a decrease in gene expression associated with cholesterol synthesis.

Theme analysis of Type III periorbital dyschromia may be broadly correlated to cellular physiological responses associated with metabolism in the epidermis and metabolism and nutrition, stress and cell communication, and structure in the dermis. In particular, a theme analysis of epidermal gene expression related to Type III periorbital dyschromia revealed, generally, a decrease in gene expression associated with oxidation and reduction process. A theme analysis of dermal gene expression revealed the following general themes: an increase in gene expression associated with ion and metal transport, regulation of the MAPK signaling cascade, regulation of Jun Kinase activity, blood vessels and neuron development and a decrease in gene expression associated with cholesterol synthesis, fatty acid metabolic process, microtubule cytoskeleton organization, organelle assembly, and carbohydrate metabolic process.

Products for treating different types of periorbital dyschromia are described in U.S. Provisional Application No. 61/798,278, filed by Osorio, et al., on Mar. 15, 2013 and titled “Array of Products.” Methods of treating different types of periorbital dyschromia are described in U.S. Provisional App. No. 61/798,340, filed by Osorio et al., on Mar. 15, 2013 and titled “Methods of Treating Periorbital Dyschromia.”

The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as “40 mm” is intended to mean “about 40 mm” Additionally, properties described herein may include one or more ranges of values. It is to be understood that these ranges include every value within the range, even though the individual values in the range may not be expressly disclosed.

All documents cited in the Detailed Description of the Invention are, in relevant part, incorporated herein by reference; the citation of any document is not to be construed as an admission that it is prior art with respect to the present invention. To the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.

While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention. 

What is claimed is:
 1. A method of identifying periorbital dyschromia, comprising: (a) identifying skin in a periorbital region of a person comprising periorbital dyschromia; (b) capturing an image of the identified skin in the periorbital region with an image capture device; (c) analyzing the image with a computer, wherein at least a portion of the image is color corrected and a measured imaging value is generated from the color corrected portion, and the measured imaging value is normalized relative to an imaging value obtained from a cheek; (d) identifying the person as exhibiting Type I periorbital dyschromia when the portion of skin is disposed in Zone 1, Zone 2 and Zone 3 of the periorbital region and the measured imaging value corresponds to at least one value from Table 1, identifying the person as exhibiting Type II periorbital dyschromia when the portion of skin is disposed in only in Zone 1 of the periorbital region and the measured imaging value corresponds to at least one value from Table 2, or identifying the person as exhibiting Type III periorbital dyschromia when the portion of skin is disposed in Zone 1 and Zone 2 and the measured imaging value corresponds to at least one value from Table 3; and (e) communicating the type of periorbital dyschromia identified in (d) to the person.
 2. The method of claim 1, wherein the measured imaging values includes at least one RGB color imaging value.
 3. The method of claim 1, wherein the imaging values are measured using an RGB imaging technique. 